Compositions and methods for the diagnosis and threatment of herpes simplex virus infection

ABSTRACT

Compounds and methods for the diagnosis and treatment of HSV infection are provided. The compounds comprise polypeptides that contain at least one antigenic portion of an HSV polypeptide and DNA sequences encoding such polypeptides. Pharmaceutical compositions and vaccines comprising such polypeptides or DNA sequences are also provided, together with antibodies directed against such polypeptides. Diagnostic kits are also provided comprising such polypeptides and/or DNA sequences and a suitable detection reagent for the detection of HSV infection in patients and in biological samples.

BACKGROUND OF THE INVENTION

[0001] 1. Field of the Invention

[0002] The present invention relates generally to the detection and treatment of HSV infection. In particular, the invention relates to polypeptides comprising HSV antigens, DNA encoding HSV antigens, and the use of such compositions for the diagnosis and treatment of HSV infection.

[0003] 2. Description of the Related Art

[0004] The herpes viruses include the herpes simplex viruses (HSV), comprising two closely related variants designated types 1 (HSV-1) and 2 (HSV-2). HSV is a prevalent cause of genital infection in humans, with an estimated annual incidence of 600,000 new cases and with 10 to 20 million individuals experiencing symptomatic chronic recurrent disease. The asymptomatic subclinical infection rate may be even higher. For example, using a type-specific serological assay, 35% of an unselected population of women attending a health maintenance organization clinic in Atlanta had antibodies to HSV type 2 (HSV-2). Although continuous administration of antiviral drugs such as acyclovir ameliorates the severity of acute HSV disease and reduces the frequency and duration of recurrent episodes, such chemotherapeutic intervention does not abort the establishment of latency nor does it alter the status of the latent virus. As a consequence, the recurrent disease pattern is rapidly reestablished upon cessation of drug treatment.

[0005] The genome of at least one strain of herpes simplex virus (HSV) has been characterized. It is approximately 150 kb and encodes about 85 known genes, each of which encodes a protein in the range of 50-1000 amino acids in length. Unknown, however, are the immunogenic portions, particularly immunogenic epitopes, that are capable of eliciting an effective T cell immune response to viral infection.

[0006] Thus, it is a matter of great medical and scientific need to identify immunogenic portions, preferably epitopes, of HSV polypeptides that are capable of eliciting an effective immune response to HSV infection. Such information will lead to safer and more effective prophylactic pharmaceutical compositions, e.g., vaccine compositions, to substantially prevent HSV infections, and, where infection has already occurred, therapeutic compositions to combat the disease. The present invention fulfills these and other needs.

BRIEF SUMMARY OF THE INVENTION

[0007] The present invention provides compositions and methods for the diagnosis and therapy of HSV infection. In one aspect, the present invention provides polypeptides comprising an immunogenic portion of a HSV antigen, or a variant or biological functional equivalent of such an antigen. Certain preferred portions and other variants are immunogenic, such that the ability of the portion or variant to react with antigen-specific antisera is not substantially diminished. Within certain embodiments, the polypeptide comprises an amino acid sequence encoded by a polynucleotide sequence selected from the group consisting of (a) a sequence of any one of SEQ ID NO: 1, 4, 8-9, 13, 16, 19 24, 35-38, 48-49, 52-53, 65-73, 76-89, 98-117, 118-119, 141, 144-152, 179-180 and 182-183; (b) a complement of said sequence; and (c) sequences that hybridize to a sequence of (a) or (b) under moderately stringent conditions. In specific embodiments, the polypeptides of the present invention comprise at least a portion, preferably at least an immunogenic portion, of a HSV protein that comprises some or all of an amino acid sequence recited in any one of SEQ ID NO: 2, 3, 5, 6, 7, 10-12, 14-15, 17-18, 20-23, 25-33, 39-47, 50-51, 54-64, 74-75, 90-97, 120-121, 122-140, 142-143, 153-178, and 181 including variants and biological functional equivalents thereof.

[0008] The present invention further provides polynucleotides that encode a polypeptide as described above, or a portion thereof (such as a portion encoding at least 15 contiguous amino acid residues of a HSV protein), expression vectors comprising such polynucleotides and host cells transformed or transfected with such expression vectors.

[0009] In a related aspect, polynucleotide sequences encoding the above polypeptides, recombinant expression vectors comprising one or more of these polynucleotide sequences and host cells transformed or transfected with such expression vectors are also provided.

[0010] In another aspect, the present invention provides fusion proteins comprising one or more HSV polypeptides, for example in combination with a physiologically acceptable carrier or immunostimulant for use as pharmaceutical compositions and vaccines thereof.

[0011] The present invention further provides pharmaceutical compositions that comprise: (a) an antibody, either polyclonal and monoclonal, or antigen-binding fragment thereof that specifically binds to a HSV protein; and (b) a physiologically acceptable carrier.

[0012] Within other aspects, the present invention provides pharmaceutical compositions that comprise one or more HSV polypeptides or portions thereof disclosed herein, or a polynucleotide molecule encoding such a polypeptide, and a physiologically acceptable carrier. The invention also provides vaccines for prophylactic and therapeutic purposes comprising one or more of the disclosed polypeptides and an immunostimulant, as defined herein, as well as vaccines comprising one or more polynucleotide sequences encoding such polypeptides and an immunostimulant.

[0013] In yet another aspect, methods are provided for inducing protective immunity in a patient, comprising administering to a patient an effective amount of one or more of the above pharmaceutical compositions or vaccines. Any of the polypeptides identified for use in the treatment of patients can be used in conjunction with pharmaceutical agents used to treat herpes infections, such as, but not limited to, Zovirax®(Acyclovir), Valtrex® (Valacyclovir), and Famvir® (Famcyclovir).

[0014] In yet a further aspect, there are provided methods for treating, substantially preventing or otherwise ameliorating the effects of an HSV infection in a patient, the methods comprising obtaining peripheral blood mononuclear cells (PBMC) from the patient, incubating the PBMC with a polypeptide of the present invention (or a polynucleotide that encodes such a polypeptide) to provide incubated T cells and administering the incubated T cells to the patient. The present invention additionally provides methods for the treatment of HSV infection that comprise incubating antigen presenting cells with a polypeptide of the present invention (or a polynucleotide that encodes such a polypeptide) to provide incubated antigen presenting cells and administering the incubated antigen presenting cells to the patient. Proliferated cells may, but need not, be cloned prior to administration to the patient. In certain embodiments, the antigen presenting cells are selected from the group consisting of dendritic cells, macrophages, monocytes, B-cells, and fibroblasts. Compositions for the treatment of HSV infection comprising T cells or antigen presenting cells that have been incubated with a polypeptide or polynucleotide of the present invention are also provided. Within related aspects, vaccines are provided that comprise: (a) an antigen presenting cell that expresses a polypeptide as described above and (b) an immunostimulant.

[0015] The present invention further provides, within other aspects, methods for removing HSV-infected cells from a biological sample, comprising contacting a biological sample with T cells that specifically react with a HSV protein, wherein the step of contacting is performed under conditions and for a time sufficient to permit the removal of cells expressing the protein from the sample.

[0016] Within related aspects, methods are provided for inhibiting the development of HSV infection in a patient, comprising administering to a patient a biological sample treated as described above. In further aspects of the subject invention, methods and diagnostic kits are provided for detecting HSV infection in a patient. In one embodiment, the method comprises: (a) contacting a biological sample with at least one of the polypeptides or fusion proteins disclosed herein; and (b) detecting in the sample the presence of binding agents that bind to the polypeptide or fusion protein, thereby detecting HSV infection in the biological sample. Suitable biological samples include whole blood, sputum, serum, plasma, saliva, cerebrospinal fluid and urine. In one embodiment, the diagnostic kits comprise one or more of the polypeptides or fusion proteins disclosed herein in combination with a detection reagent. In yet another embodiment, the diagnostic kits comprise either a monoclonal antibody or a polyclonal antibody that binds with a polypeptide of the present invention.

[0017] The present invention also provides methods for detecting HSV infection comprising: (a) obtaining a biological sample from a patient; (b) contacting the sample with at least two oligonucleotide primers in a polymerase chain reaction, at least one of the oligonucleotide primers being specific for a polynucleotide sequence disclosed herein; and (c) detecting in the sample a polynucleotide sequence that amplifies in the presence of the oligonucleotide primers. In one embodiment, the oligonucleotide primer comprises at about 10 contiguous nucleotides of a polynucleotide sequence peptide disclosed herein, or of a sequence that hybridizes thereto.

[0018] In a further aspect, the present invention provides a method for detecting HSV infection in a patient comprising: (a) obtaining a biological sample from the patient; (b) contacting the sample with an oligonucleotide probe specific for a polynucleotide sequence disclosed herein; and (c) detecting in the sample a polynucleotide sequence that hybridizes to the oligonucleotide probe. In one embodiment, the oligonucleotide probe comprises at least about 15 contiguous nucleotides of a polynucleotide sequence disclosed herein, or a sequence that hybridizes thereto.

[0019] These and other aspects of the present invention will become apparent upon reference to the following detailed description. All references disclosed herein are hereby incorporated by reference in their entirety as if each was incorporated individually.

BRIEF DESCRIPTION OF THE SEVERAL SEQUENCE IDENTIFIERS

[0020] SEQ ID NO: 1 sets forth a polynucleotide sequence of an isolated clone designated HSV2I_UL39frag12A12;

[0021] SEQ ID NO: 2 sets forth an amino acid sequence, designated H12A12orf1.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 1;

[0022] SEQ ID NO: 3 sets forth the amino acid sequence of the full length HSV-2 UL39 protein;

[0023] SEQ ID NO: 4 sets forth a polynucleotide sequence of an isolated clone designated HSV2II_US8AfragD6.B_B11_T7Trc.seq;

[0024] SEQ ID NO: 5 sets forth an amino acid sequence, designated D6Borf1.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 4;

[0025] SEQ ID NO: 6 sets forth an amino acid sequence, designated D6Borf2.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 4;

[0026] SEQ ID NO: 7 sets forth the amino acid sequence of the full length HSV-2 US8A protein;

[0027] SEQ ID NO: 8 sets forth a polynucleotide sequence of an isolated clone designated HSV2II_US4fragF10B3_T7Trc.seq;

[0028] SEQ ID NO: 9 sets forth a polynucleotide sequence of an isolated clone designated HSV2II_US3fragF10B3_T7P.seq;

[0029] SEQ ID NO: 10 sets forth an amino acid sequence, designated F10B3orf2.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO:8;

[0030] SEQ ID NO: 11 sets forth an amino acid sequence, designated 8F10B3orf1.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 9;

[0031] SEQ ID NO: 12 sets forth the amino acid sequence of the full length HSV-2 US3 protein;

[0032] SEQ ID NO: 13 sets forth a polynucleotide sequence of an isolated clone designated HSV2II_UL46fragF11F5_T7Trc.seq

[0033] SEQ ID NO: 14 sets forth an amino acid sequence, designated F11F5orf1.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 13;

[0034] SEQ ID NO: 15 sets forth the amino acid sequence of the full length HSV-2 UL46 protein;

[0035] SEQ ID NO: 16 sets forth a polynucleotide sequence of an isolated clone designated HSV2II_UL27fragH2C7_T7Trc.seq

[0036] SEQ ID NO: 17 sets forth an amino acid sequence, designated H2C7orf1.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 16;

[0037] SEQ ID NO: 18 sets forth the amino acid sequence of the full length HSV-2 UL27 protein;

[0038] SEQ ID NO: 19 sets forth a polynucleotide sequence of an isolated clone designated HSV2II_UL18fragF10A1_rc.seq;

[0039] SEQ ID NO: 20 sets forth an amino acid sequence, designated F10A1 orf3.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 19;

[0040] SEQ ID NO: 21 sets forth an amino acid sequence, designated F10A1 orf2.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 19;

[0041] SEQ ID NO: 22 sets forth an amino acid sequence, designated F10A1orf1.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 19;

[0042] SEQ ID NO: 23 sets forth the amino acid sequence of the full length HSV-2 UL18 protein;

[0043] SEQ ID NO: 24 sets forth a polynucleotide sequence of an isolated clone designated HSV2II_UL15fragF10A12_rc.seq;

[0044] SEQ ID NO: 25 sets forth an amino acid sequence, designated F10A12orf1.pro, of an open reading frame encoded within the polynucleotide of SEQ ID NO: 24;

[0045] SEQ ID NO: 26 sets forth the amino acid sequence of the full length HSV-2 UL15 protein;

[0046] SEQ ID NO:27 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL46 gene;

[0047] SEQ ID NO:28 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL46 gene;

[0048] SEQ ID NO:29 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL46 gene;

[0049] SEQ ID NO:30 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL46 gene;

[0050] SEQ ID NO:31 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL46 gene;

[0051] SEQ ID NO:32 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL18 gene;

[0052] SEQ ID NO:33 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL18 gene;

[0053] SEQ ID NO:34 sets forth a nucleotide sequence of an isolated clone designated RL2_E9A4_(—)5_consensus.seq;

[0054] SEQ ID NO:35 sets forth the nucleotide sequence of the full length HSV-2 RL2 gene;

[0055] SEQ ID NO:36 sets for the nucleotide sequence of an isolated clone designated UL23_(—)22_C12A12_consensus.seq;

[0056] SEQ ID NO:37 sets forth the nucleotide sequence of the full length HSV-2 UL23 protein;

[0057] SEQ ID NO:38 sets forth the nucleotide sequence of the full length HSV-2 UL22 protein;

[0058] SEQ ID NO:39 sets forth an amino acid sequence, designated HSV2_UL23, of an open reading frame encoded by the polynucleotide of SEQ ID NO: 37;

[0059] SEQ ID NO:40 sets forth an amino acid sequence designated HSV2_UL23 of an open reading frame encoded within the polynucleotides of SEQ ID NO:36;

[0060] SEQ ID NO:41 sets forth an amino acid sequence designated HSV2_UL22 of an open reading frame encoded within the polynucleotides of SEQ ID NO:36;

[0061] SEQ ID NO:42 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL23 gene;

[0062] SEQ ID NO:43 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL23 gene;

[0063] SEQ ID NO:44 sets forth the amino acid sequence of a 15-mer polypeptide derived from an immunogenic portion of the HSVII UL23 gene;

[0064] SEQ ID NO:45 sets forth an amino acid sequence, designated HSV2_UL22, of an open reading frame encoded by the polynucleotide of SEQ ID NO:38;

[0065] SEQ ID NO:46 sets forth an amino acid sequence, designated RL2_E9A4_(—)5_consensus.seq, of an open reading frame encoded by the polynucleotide of SEQ ID NO:34;

[0066] SEQ ID NO:47 sets forth an amino acid sequence, designated HSV2_RL2, of an open reading frame encoded by the polynucleotide of SEQ ID NO:35;

[0067] SEQ ID NO:48 sets forth a nucleotide sequence of an isolated clone designated G10_UL37consensus.seq;

[0068] SEQ ID NO:49 sets forth the nucleotide sequence of the full length HSV-2 UL37 gene;

[0069] SEQ ID NO:50 sets forth an amino acid sequence, designated HSV2_UL37, of an open reading frame encoded by the polynucleotide of SEQ ID NO:48; and

[0070] SEQ ID NO:51 sets forth an amino acid sequence, designated HSV2_UL37, of an open reading frame encoded by the polynucleotide of SEQ ID NO:49;

[0071] SEQ ID NO:52 sets forth the DNA sequence derived from the insert of clone UL46fragF11 F5;

[0072] SEQ ID NO:53 sets forth the DNA sequence derived from the insert of clone G10;

[0073] SEQ ID NO:54 sets forth the amino acid sequence derived from the insert of clone UL46fragF11F5;

[0074] SEQ ID NO:55 sets forth the amino acid sequence derived from the insert of clone G10;

[0075] SEQ ID NO:56 is amino acid sequence of peptide #23 (amino acids 688-702) of the HSV-2 gene UL15;

[0076] SEQ ID NO:57 is amino acid sequence of peptide #30 (amino acids 716-730) of the HSV-2 gene UL15;

[0077] SEQ ID NO:58 is amino acid sequence of peptide #7 (amino acids 265-279) of the HSV-2 gene UL23;

[0078] SEQ ID NO:59 is amino acid sequence of peptide #2 (amino acids 621-635) of the HSV-2 gene UL46;

[0079] SEQ ID NO:60 is amino acid sequence of peptide #8 (amino acids 645-659) of the HSV-2 gene UL46;

[0080] SEQ ID NO:61 is amino acid sequence of peptide #9 (amino acids 649-663) of the HSV-2 gene UL46;

[0081] SEQ ID NO:62 is amino acid sequence of peptide #11 (amino acids 657-671) of the HSV-2 gene UL46;

[0082] SEQ ID NO:63 is amino acid sequence of peptide #33 (amino acids 262-276) of the HSV-2 gene US3;

[0083] SEQ ID NO:64 is amino acid sequence of peptide #5 (amino acids 99-113) of the HSV-2 gene US8A.

[0084] SEQ ID NO:65 sets forth the polynucleotide sequence of the full length HSV-2 UL39 protein.

[0085] SEQ ID NO:66 sets forth the partial polynucleotide sequence of UL39 derived from the HSV2-III library, pools 1F4, 1G2, and 3G11 which were recognized by clone 39.

[0086] SEQ ID NO:67 sets forth the partial polynucleotide sequence of UL39 derived from the HSV2-III library, pool 2C4 which was recognized by clone 39.

[0087] SEQ ID NO:68 sets forth the 5′ end of the partial polynucleotide sequence of ICP0 derived from the HSV2-III library, pools 3H6, 3F12, and 4B2 which were recognized by clone 47.

[0088] SEQ ID NO:69 sets forth the 3′ end of the partial polynucleotide sequence of ICP0 derived from the HSV2-III library, pools 3H6, 3F12, and 4B2 which were recognized by clone 47.

[0089] SEQ ID NO:70 sets forth the 5′ end of the partial polynucleotide sequence of ICP0 derived from the HSV2-III library, pool 3A1 which was recognized by clone 47.

[0090] SEQ ID NO:71 sets forth the 3′ end of the partial polynucleotide sequence of ICP0 derived from the HSV2-III library, pool 3A1 which was recognized by clone 47.

[0091] SEQ ID NO:72 sets forth the 5′ end of the partial polynucleotide sequence of ICP0 derived from the HSV2-III library, pool 2B2 which was recognized by clone 47.

[0092] SEQ ID NO:73 sets forth the 3′ end of the partial polynucleotide sequence of ICP0 derived from the HSV2-III library, pool 2B2 which was recognized by clone 47.

[0093] SEQ ID NO:74 sets forth the partial amino acid sequence of UL39 derived from the HSV2-III library, pools 1F4, 1G2, and 3G11 which were recognized by clone 39.

[0094] SEQ ID NO:75 sets forth the partial amino acid sequence of UL39 derived from the HSV2-III library, pool 2C4 which was recognized by clone 39.

[0095] SEQ ID NO:76 sets forth a full length DNA sequence for the HSV-2 gene UL19.

[0096] SEQ ID NO:77 sets forth a DNA sequence for the vaccinia virus shuttle plasmid, pSC11.

[0097] SEQ ID NO:78 sets forth a full length DNA sequence for the HSV-2 gene, UL47.

[0098] SEQ ID NO:79 sets forth a full length DNA sequence for the HSV-2 gene, UL50.

[0099] SEQ ID NO:80 sets forth a DNA sequence for the human Ubiquitin gene.

[0100] SEQ ID NO:81 sets forth a full length DNA sequence for the HSV-2 gene, UL49.

[0101] SEQ ID NO:82 sets forth a DNA sequence corresponding to the coding region of the HSV gene, UL50.

[0102] SEQ ID NO:83 sets forth a DNA sequence corresponding to the coding region of the HSV gene, UL49.

[0103] SEQ ID NO:84 sets forth a DNA sequence corresponding to the coding region of the HSV gene, UL19.

[0104] SEQ ID NO:85 sets forth a DNA sequence corresponding to the coding region of the HSV gene, UL21.

[0105] SEQ ID NO:86 sets forth a DNA sequence corresponding to the coding region of the HSV-2 UL47 gene with the Trx2 fusion sequence.

[0106] SEQ ID NO:87 sets forth a DNA sequence corresponding to the coding region of the HSV gene, UL47.

[0107] SEQ ID NO:88 sets forth a DNA sequence corresponding to the coding region of the HSV gene, UL47 C fragment.

[0108] SEQ ID NO:89 sets forth a DNA sequence corresponding to the coding region of the HSV gene, UL39.

[0109] SEQ ID NO:90 sets forth an amino acid sequence corresponding to the UL39 protein with a His tag.

[0110] SEQ ID NO:91 sets forth an amino acid sequence corresponding to the UL21 protein with a His tag.

[0111] SEQ ID NO:92 sets forth an amino acid sequence corresponding to the UL47 protein fused with the Trx and 2 histadine tags.

[0112] SEQ ID NO:93 sets forth an amino acid sequence corresponding to the UL47 C fragment with a His tag.

[0113] SEQ ID NO:94 sets forth an amino acid sequence corresponding to the UL47 protein with a His tag.

[0114] SEQ ID NO:95 sets forth an amino acid sequence corresponding to the UL19 protein with a His tag.

[0115] SEQ ID NO:96 sets forth an amino acid sequence corresponding to the UL50 protein with a His tag.

[0116] SEQ ID NO:97 sets forth an amino acid sequence corresponding to the UL49 protein with a His tag.

[0117] SEQ ID NO:98 sets forth the primer sequence for the sense primer PDM-602, used in the amplification of UL21.

[0118] SEQ ID NO:99 sets forth the primer sequence for the reverse primer PDM-603, used in the amplification of UL21.

[0119] SEQ ID NO:100 sets forth the primer sequence for the sense primer PDM-466, used in the amplification of UL39.

[0120] SEQ ID NO:101 sets forth the primer sequence for the reverse primer PDM-467, used in the amplification of UL39.

[0121] SEQ ID NO:102 sets forth the primer sequence for the sense primer PDM-714, used in the amplification of UL49.

[0122] SEQ ID NO:103 sets forth the primer sequence for the reverse primer PDM-715, used in the amplification of UL49.

[0123] SEQ ID NO: 104 sets forth the primer sequence for the sense primer PDM-458, used in the amplification of UL50.

[0124] SEQ ID NO:105 sets forth the primer sequence for the reverse primer PDM-459, used in the amplification of UL50.

[0125] SEQ ID NO:106 sets forth the primer sequence for the sense primer PDM-453, used in the amplification of UL19.

[0126] SEQ ID NO:107 sets forth the primer sequence for the reverse primer PDM-457, used in the amplification of UL19.

[0127] SEQ ID NO:108 sets forth the primer sequence for the sense primer PDM-631, used in the amplification of UL47.

[0128] SEQ ID NO: 109 sets forth the primer sequence for the reverse primer PDM-632, used in the amplification of UL47.

[0129] SEQ ID NO:110 sets forth the primer sequence for the sense primer PDM-631, used in the amplification of UL47 A.

[0130] SEQ ID NO:111 sets forth the primer sequence for the reverse primer PDM-645, used in the amplification of UL47 A.

[0131] SEQ ID NO:112 sets forth the primer sequence for the sense primer PDM-646, used in the amplification of UL47 B.

[0132] SEQ ID NO:113 sets forth the primer sequence for the reverse primer PDM-632, used in the amplification of UL47 B.

[0133] SEQ ID NO:114 sets forth the primer sequence for the sense primer PDM-631, used in the amplification of UL47 C.

[0134] SEQ ID NO:115 sets forth the primer sequence for the reverse primer PDM-739, used in the amplification of UL47 C.

[0135] SEQ ID NO:116 sets forth the primer sequence for the sense primer PDM-740, used in the amplification of UL47 D.

[0136] SEQ ID NO:117 sets forth the primer sequence for the reverse primer PDM-632, used in the amplification of UL47 D.

[0137] SEQ ID NO:118 sets forth a novel DNA sequence for the HSV-2 gene, US8.

[0138] SEQ ID NO:119 sets forth the published DNA sequence for the HSV-2 gene, US8, derived from the HG52 strain of HSV-2.

[0139] SEQ ID NO:120 sets forth an amino acid sequence encoded by SEQ ID NO:118.

[0140] SEQ ID NO:121 sets forth an amino acid sequence encoded by SEQ ID NO:119.

[0141] SEQ ID NO:122 sets forth the sequence of peptide 85 (p85), a CD8+ peptide derived from the HSV-2 gene, UL47.

[0142] SEQ ID NO:123 sets forth the sequence of peptide 89 (p89), a CD8+ peptide derived from the HSV-2 gene, UL47.

[0143] SEQ ID NO:124 sets forth the sequence of peptide 98/99 (p98/99), a CD8+ peptide derived from the HSV-2 gene, UL47.

[0144] SEQ ID NO:125 sets forth the sequence of peptide 105 (p105), a CD8+ peptide derived from the HSV-2 gene, UL47.

[0145] SEQ ID NO:126 sets forth the sequence of peptide 112 (p112), a CD8+ peptide derived from the HSV-2 gene, UL47.

[0146] SEQ ID NO:127 sets forth the sequence of peptide #23 (amino acids 688-702) from the HSV-2 protein UL15.

[0147] SEQ ID NO:128 sets forth the sequence of peptide #30 (amino acids 716-730) from the HSV-2 protein UL15.

[0148] SEQ ID NO:129 sets forth the sequence of peptide #7 (amino acids 265-272) from the HSV-2 protein UL23.

[0149] SEQ ID NO:130 sets forth the sequence of peptide #2 (amino acids 621-635) from the HSV-2 protein UL46.

[0150] SEQ ID NO:131 sets forth the sequence of peptide #8 (amino acids 645-659) from the HSV-2 protein UL46.

[0151] SEQ ID NO:132 sets forth the sequence of peptide #9 (amino acids 649-663) from the HSV-2 protein UL46.

[0152] SEQ ID NO:133 sets forth the sequence of peptide #11 (amino acids 657-671) from the HSV-2 protein UL46.

[0153] SEQ ID NO:134 sets forth the sequence of peptide #86 (amino acids 341-355) from the HSV-2 protein UL47.

[0154] SEQ ID NO:135 sets forth the sequence of peptide #6 (amino acids 21-35) from the HSV-2 protein UL49.

[0155] SEQ ID NO:136 sets forth the sequence of peptide #12 (amino acids 45-59) from the HSV-2 protein UL49.

[0156] SEQ ID NO:137 sets forth the sequence of peptide #13 (amino acids 49-63) from the HSV-2 protein UL49.

[0157] SEQ ID NO:138 sets forth the sequence of peptide #49 (amino acids 193-208) from the HSV-2 protein UL49.

[0158] SEQ ID NO:139 sets forth the sequence of peptide #33 (amino acids 262-276) from the HSV-2 protein US3.

[0159] SEQ ID NO: 140 sets forth the sequence of peptide #5 (amino acids 99-113) from the HSV-2 protein US8A.

[0160] SEQ ID NO:141 sets forth a full length insert DNA sequence corresponding to the clone F10B3.

[0161] SEQ ID NO:142 sets forth a full length insert amino acid sequence corresponding to the clone F10B3.

[0162] SEQ ID NO:143 sets forth an amino acid sequence for the HSV-2 protein, US4.

[0163] SEQ ID NO:144 sets forth a DNA sequence for the HSV-2 protein, UL21.

[0164] SEQ ID NO:145 sets forth a DNA sequence for the HSV-2 protein, UL50.

[0165] SEQ ID NO:146 sets forth a DNA sequence for the HSV-2 protein, US3.

[0166] SEQ ID NO:147 sets forth a DNA sequence for the HSV-2 protein, UL54.

[0167] SEQ ID NO:148 sets forth a DNA sequence for the HSV-2 protein, US8.

[0168] SEQ ID NO:149 sets forth a DNA sequence for the HSV-2 protein, UL19.

[0169] SEQ ID NO:150 sets forth a DNA sequence for the HSV-2 protein, UL46.

[0170] SEQ ID NO:151 sets forth a DNA sequence for the HSV-2 protein, UL18.

[0171] SEQ ID NO:152 sets forth a DNA sequence for the HSV-2 protein, RL2.

[0172] SEQ ID NO:153 sets forth an amino sequence for the HSV-2 protein, UL50.

[0173] SEQ ID NO:154 sets forth an amino acid sequence for the HSV-2 protein, UL21.

[0174] SEQ ID NO:155 sets forth an amino acid sequence for the HSV-2 protein, US3.

[0175] SEQ ID NO:156 sets forth an amino acid sequence for the HSV-2 protein, UL54.

[0176] SEQ ID NO:157 sets forth an amino acid sequence for the HSV-2 protein, US8.

[0177] SEQ ID NO:158 sets forth an amino acid sequence for the HSV-2 protein, UL19.

[0178] SEQ ID NO:159 sets forth an amino acid sequence for the HSV-2 protein, UL46.

[0179] SEQ ID NO:160 sets forth an amino acid sequence for the HSV-2 protein, UL18.

[0180] SEQ ID NO:161 sets forth an amino acid sequence for the HSV-2 protein, RL2.

[0181] SEQ ID NO:162 sets forth the sequence of peptide #43 (amino acids 211-225) from the HSV-2 protein RL2.

[0182] SEQ ID NO:163 sets forth the sequence of peptide #41 (amino acids 201-215) from the HSV-2 protein UL46.

[0183] SEQ ID NO:164 sets forth the sequence of peptide #50 (amino acids 246-260) from the HSV-2 protein UL46.

[0184] SEQ ID NO:165 sets forth the sequence of peptide #51 (amino acids 251-265) from the HSV-2 protein UL46.

[0185] SEQ ID NO:166 sets forth the sequence of peptide #60 (amino acids 296-310) from the HSV-2 protein UL46.

[0186] SEQ ID NO:167 sets forth the sequence of peptide #74 (amino acids 366-380) from the HSV-2 protein US8.

[0187] SEQ ID NO:168 sets forth the sequence of peptide #102 (amino acids 506-520) from the HSV-2 protein UL19.

[0188] SEQ ID NO:169 sets forth the sequence of peptide #103 (amino acids 511-525) from the HSV-2 protein UL19.

[0189] SEQ ID NO:170 sets forth the sequence of peptide #74 (amino acids 366-380) from the HSV-2 protein UL19.

[0190] SEQ ID NO:171 sets forth the sequence of peptide #75 (amino acids 371-385) from the HSV-2 protein UL19.

[0191] SEQ ID NO:172 sets forth the sequence of peptide #17 (amino acids 65-79) from the HSV-2 protein UL18.

[0192] SEQ ID NO:173 sets forth the sequence of peptide #18 (amino acids 69-83) from the HSV-2 protein UL18.

[0193] SEQ ID NO:174 sets forth the sequence of peptide #16 (amino acids 76-90) from the HSV-2 protein UL50.

[0194] SEQ ID NO:175 sets forth the sequence of peptide #23 (amino acids 111-125) from the HSV-2 protein UL50.

[0195] SEQ ID NO:176 sets forth the sequence of peptide #49 (amino acids 241-255) from the HSV-2 protein UL50.

[0196] SEQ ID NO:177 sets forth the sequence of a 9-mer peptide for ICP0 (amino acids 215-223).

[0197] SEQ ID NO:178 sets forth the sequence of a 10-mer peptide for UL46 (amino acids 251-260).

[0198] SEQ ID NO:179 sets forth a DNA sequence of US4 derived from the HG52 strain of HSV-2.

[0199] SEQ ID NO:180 sets forth a DNA sequence for the UL47 F coding region.

[0200] SEQ ID NO:181 sets forth an amino acid sequence for the UL47 F coding region.

[0201] SEQ ID NO:182 sets forth the sequence for primer CBH-002 used in the amplification of UL47 F.

[0202] SEQ ID NO:183 sets forth the sequence for primer PDM-632 used in the amplification of UL47 F.

DETAILED DESCRIPTION OF THE INVENTION

[0203] U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification and/or listed in the Application Data Sheet, are incorporated herein by reference, in their entirety.

[0204] As noted above, the present invention is generally directed to compositions and methods for making and using the compositions, particularly in the therapy and diagnosis of HSV infection. Certain illustrative compositions described herein include HSV polypeptides, polynucleotides encoding such polypeptides, binding agents such as antibodies, antigen presenting cells (APCs) and/or immune system cells (e.g., T cells). Certain HSV proteins and immunogenic portions thereof comprise HSV polypeptides that react detectably (within an immunoassay, such as an ELISA or Western blot) with antisera of a patient infected with HSV.

[0205] Therefore, the present invention provides illustrative polynucleotide compositions, illustrative polypeptide compositions, immunogenic portions of said polynucleotide and polypeptide compositions, antibody compositions capable of binding such polypeptides, and numerous additional embodiments employing such compositions, for example in the detection, diagnosis and/or therapy of human HSV infections.

[0206] Polynucleotide Compositions

[0207] As used herein, the terms “DNA segment” and “polynucleotide” refer to a DNA molecule that has been isolated free of total genomic DNA of a particular species. Therefore, a DNA segment encoding a polypeptide refers to a DNA segment that contains one or more coding sequences yet is substantially isolated away from, or purified free from, total genomic DNA of the species from which the DNA segment is obtained. Included within the terms “DNA segment” and “polynucleotide” are DNA segments and smaller fragments of such segments, and also recombinant vectors, including, for example, plasmids, cosmids, phagemids, phage, viruses, and the like.

[0208] As will be understood by those skilled in the art, the DNA segments of this invention can include genomic sequences, extra-genomic and plasmid-encoded sequences and smaller engineered gene segments that express, or may be adapted to express, proteins, polypeptides, peptides and the like. Such segments may be naturally isolated, or modified synthetically by the hand of man.

[0209] “Isolated,” as used herein, means that a polynucleotide is substantially away from other coding sequences, and that the DNA segment does not contain large portions of unrelated coding DNA, such as large chromosomal fragments or other functional genes or polypeptide coding regions. Of course, this refers to the DNA segment as originally isolated, and does not exclude genes or coding regions later added to the segment by the hand of man.

[0210] As will be recognized by the skilled artisan, polynucleotides may be single-stranded (coding or antisense) or double-stranded, and may be DNA (genomic, cDNA or synthetic) or RNA molecules. RNA molecules include HnRNA molecules, which contain introns and correspond to a DNA molecule in a one-to-one manner, and mRNA molecules, which do not contain introns. Additional coding or non-coding sequences may, but need not, be present within a polynucleotide of the present invention, and a polynucleotide may, but need not, be linked to other molecules and/or support materials.

[0211] Polynucleotides may comprise a native sequence (i.e., an endogenous sequence that encodes an HSV protein or a portion thereof) or may comprise a variant, or a biological or antigenic functional equivalent of such a sequence. Polynucleotide variants may contain one or more substitutions, additions, deletions and/or insertions, as further described below, preferably such that the immunogenicity of the encoded polypeptide is not diminished, relative to a native HSV protein. The effect on the immunogenicity of the encoded polypeptide may generally be assessed as described herein. The term “variants” also encompasses homologous genes of xenogenic origin.

[0212] When comparing polynucleotide or polypeptide sequences, two sequences are said to be “identical” if the sequence of nucleotides or amino acids in the two sequences is the same when aligned for maximum correspondence, as described below. Comparisons between two sequences are typically performed by comparing the sequences over a comparison window to identify and compare local regions of sequence similarity. A “comparison window” as used herein, refers to a segment of at least about 20 contiguous positions, usually 30 to about 75, 40 to about 50, in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned.

[0213] Optimal alignment of sequences for comparison may be conducted using the Megalign program in the Lasergene suite of bioinformatics software (DNASTAR, Inc., Madison, Wis.), using default parameters. This program embodies several alignment schemes described in the following references: Dayhoff, M. O., A model of evolutionary change in proteins—Matrices for detecting distant relationships, 1978. In Dayhoff, M. O. (ed.), Atlas of Protein Sequence and Structure, National Biomedical Research Foundation, Washington D.C., Vol. 5, Suppl. 3, pp. 345-358; Hein J., Unified Approach to Alignment and Phylogenes, “Methods in Enzymology,” Academic Press, Inc., San Diego, CAvol. 183, pp. 626-645,1990; Higgins, D. G. and P. M. Sharp, CABIOS 5:151-53,1989; Myers, E. W. and W. Muller, CABIOS 4:11-17,1988; Robinson, E. D., Comb. Theor 11:105, 1971; Santou, N. and M. Nes, Mol. Biol. Evol. 4:406-25,1987; Sneath, P. H. A. and R. R. Sokal, Numerical Taxonomy—the Principles and Practice of Numerical Taxonomy, Freeman Press, San Francisco, Calif., 1973; Wilbur, W. J. and D. J. Lipman, Proc. Natl. Acad., Sci. USA 80:726-30,1983.

[0214] Alternatively, optimal alignment of sequences for comparison may be conducted by the local identity algorithm of Smith and Waterman, Add. APL. Math 2:482,1981, by the identity alignment algorithm of Needleman and Wunsch, J. Mol. Biol. 48:443,1970, by the search for similarity methods of Pearson and Lipman, Proc. Natl. Acad. Sci. USA 85:2444,1988, by computerized implementations of these algorithms (GAP, BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Dr., Madison, Wis.), or by inspection.

[0215] One preferred example of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nucl. Acids Res. 25:3389-3402,1977; and Altschul et al., J. Mol. Biol. 215:403-10,1990, respectively. BLAST and BLAST 2.0 can be used, for example with the parameters described herein, to determine percent sequence identity for the polynucleotides and polypeptides of the invention. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. In one illustrative example, cumulative scores can be calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix can be used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915, 1989) alignments, (B) of 50, expectation (E) of 10, M=5, N=−4 and a comparison of both strands.

[0216] Preferably, the “percentage of sequence identity” is determined by comparing two optimally aligned sequences over a window of comparison of at least 20 positions, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) of 20 percent or less, usually 5 to 15 percent, or 10 to 12 percent, as compared to the reference sequences (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid bases or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the reference sequence (i.e., the window size) and multiplying the results by 100 to yield the percentage of sequence identity.

[0217] Therefore, the present invention encompasses polynucleotide and polypeptide sequences having substantial identity to the sequences disclosed herein, for example those comprising at least 50% sequence identity, preferably at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% or higher, sequence identity compared to a polynucleotide or polypeptide sequence of this invention using the methods described herein, (e.g., BLAST analysis using standard parameters, as described below). One skilled in this art will recognize that these values can be appropriately adjusted to determine corresponding identity of proteins encoded by two nucleotide sequences by taking into account codon degeneracy, amino acid similarity, reading frame positioning and the like.

[0218] In additional embodiments, the present invention provides isolated polynucleotides and polypeptides comprising various lengths of contiguous stretches of sequence identical to or complementary to one or more of the sequences disclosed herein. For example, polynucleotides are provided by this invention that comprise at least about 15, 20, 30, 40, 50, 75, 100, 150, 200, 300, 400, 500 or 1000 or more contiguous nucleotides of one or more of the sequences disclosed herein as well as all intermediate lengths there between. It will be readily understood that “intermediate lengths”, in this context, means any length between the quoted values, such as 16, 17, 18, 19, etc.; 21, 22, 23, etc.; 30, 31, 32, etc.; 50, 51, 52, 53, etc.; 100, 101, 102, 103, etc.; 150, 151, 152, 153, etc.; including all integers through 200-500; 500-1,000, and the like.

[0219] The polynucleotides of the present invention, or fragments thereof, regardless of the length of the coding sequence itself, may be combined with other DNA sequences, such as promoters, polyadenylation signals, additional restriction enzyme sites, multiple cloning sites, other coding segments, and the like, such that their overall length may vary considerably. It is therefore contemplated that a nucleic acid fragment of almost any length may be employed, with the total length preferably being limited by the ease of preparation and use in the intended recombinant DNA protocol. For example, illustrative DNA segments with total lengths of about 10,000, about 5000, about 3000, about 2,000, about 1,000, about 500, about 200, about 100, about 50 base pairs in length, and the like, (including all intermediate lengths) are contemplated to be useful in many implementations of this invention.

[0220] In other embodiments, the present invention is directed to polynucleotides that are capable of hybridizing under moderately stringent conditions to a polynucleotide sequence provided herein, or a fragment thereof, or a complementary sequence thereof. Hybridization techniques are well known in the art of molecular biology. For purposes of illustration, suitable moderately stringent conditions for testing the hybridization of a polynucleotide of this invention with other polynucleotides include prewashing in a solution of 5× SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0); hybridizing at 50° C.-65° C., 5× SSC, overnight; followed by washing twice at 65° C. for 20 minutes with each of 2×, 0.5× and 0.2× SSC containing 0.1% SDS.

[0221] Moreover, it will be appreciated by those of ordinary skill in the art that, as a result of the degeneracy of the genetic code, there are many nucleotide sequences that encode a polypeptide as described herein. Some of these polynucleotides bear minimal homology to the nucleotide sequence of any native gene. Nonetheless, polynucleotides that vary due to differences in codon usage are specifically contemplated by the present invention. Further, alleles of the genes comprising the polynucleotide sequences provided herein are within the scope of the present invention. Alleles are endogenous genes that are altered as a result of one or more mutations, such as deletions, additions and/or substitutions of nucleotides. The resulting mRNA and protein may, but need not, have an altered structure or function. Alleles may be identified using standard techniques (such as hybridization, amplification and/or database sequence comparison).

[0222] Probes and Primers

[0223] In other embodiments of the present invention, the polynucleotide sequences provided herein can be advantageously used as probes or primers for nucleic acid hybridization. As such, it is contemplated that nucleic acid segments that comprise a sequence region of at least about 15 nucleotide long contiguous sequence that has the same sequence as, or is complementary to, a 15 nucleotide long contiguous sequence disclosed herein will find particular utility. Longer contiguous identical or complementary sequences, e.g., those of about 20, 30, 40, 50, 100, 200, 500, 1000 (including all intermediate lengths) and even up to full length sequences will also be of use in certain embodiments.

[0224] The ability of such nucleic acid probes to specifically hybridize to a sequence of interest will enable them to be of use in detecting the presence of complementary sequences in a given sample. However, other uses are also envisioned, such as the use of the sequence information for the preparation of mutant species primers, or primers for use in preparing other genetic constructions.

[0225] Polynucleotide molecules having sequence regions consisting of contiguous nucleotide stretches of 10-14, 15-20, 30, 50, or even of 100-200 nucleotides or so (including intermediate lengths as well), identical or complementary to a polynucleotide sequence disclosed herein, are particularly contemplated as hybridization probes for use in, e.g., Southern and Northern blotting. This would allow a gene product, or fragment thereof, to be analyzed, both in diverse cell types and also in various bacterial cells. The total size of fragment, as well as the size of the complementary stretch(es), will ultimately depend on the intended use or application of the particular nucleic acid segment. Smaller fragments will generally find use in hybridization embodiments, wherein the length of the contiguous complementary region may be varied, such as between about 15 and about 100 nucleotides, but larger contiguous complementarity stretches may be used, according to the length complementary sequences one wishes to detect.

[0226] The use of a hybridization probe of about 15-25 nucleotides in length allows the formation of a duplex molecule that is both stable and selective. Molecules having contiguous complementary sequences over stretches greater than 15 bases in length are generally preferred, though, in order to increase stability and selectivity of the hybrid, and thereby improve the quality and degree of specific hybrid molecules obtained. One will generally prefer to design nucleic acid molecules having gene-complementary stretches of 15 to 25 contiguous nucleotides, or even longer where desired.

[0227] Hybridization probes may be selected from any portion of any of the sequences disclosed herein. All that is required is to review the sequence set forth in the sequences disclosed herein, or to any continuous portion of the sequence, from about 15-25 nucleotides in length up to and including the full length sequence, that one wishes to utilize as a probe or primer. The choice of probe and primer sequences may be governed by various factors. For example, one may wish to employ primers from towards the termini of the total sequence.

[0228] Small polynucleotide segments or fragments may be readily prepared by, for example, directly synthesizing the fragment by chemical means, as is commonly practiced using an automated oligonucleotide synthesizer. Also, fragments may be obtained by application of nucleic acid reproduction technology, such as the PCR™ technology of U.S. Pat. No. 4,683,202 (incorporated herein by reference), by introducing selected sequences into recombinant vectors for recombinant production, and by other recombinant DNA techniques generally known to those of skill in the art of molecular biology.

[0229] The nucleotide sequences of the invention may be used for their ability to selectively form duplex molecules with complementary stretches of the entire gene or gene fragments of interest. Depending on the application envisioned, one will typically desire to employ varying conditions of hybridization to achieve varying degrees of selectivity of probe towards target sequence. For applications requiring high selectivity, one will typically desire to employ relatively stringent conditions to form the hybrids, e.g., one will select relatively low salt and/or high temperature conditions, such as provided by a salt concentration of from about 0.02 M to about 0.15 M salt at temperatures of from about 50° C. to about 70° C. Such selective conditions tolerate little, if any, mismatch between the probe and the template or target strand, and would be particularly suitable for isolating related sequences.

[0230] Of course, for some applications, for example, where one desires to prepare mutants employing a mutant primer strand hybridized to an underlying template, less stringent (reduced stringency) hybridization conditions will typically be needed in order to allow formation of the heteroduplex. In these circumstances, one may desire to employ salt conditions such as those of from about 0.15 M to about 0.9 M salt, at temperatures ranging from about 20° C. to about 55° C. Cross-hybridizing species can thereby be readily identified as positively hybridizing signals with respect to control hybridizations. In any case, it is generally appreciated that conditions can be rendered more stringent by the addition of increasing amounts of formamide, which serves to destabilize the hybrid duplex in the same manner as increased temperature. Thus, hybridization conditions can be readily manipulated, and thus will generally be a method of choice depending on the desired results.

[0231] Polynucleotide Identification and Characterization

[0232] Polynucleotides may be identified, prepared and/or manipulated using any of a variety of well established techniques. For example, a polynucleotide may be identified, as described in more detail below, by screening a microarray of cDNAs for HSV-associated expression (i.e., expression that is at least two fold greater in infected versus normal tissue, as determined using a representative assay provided herein). Such screens may be performed, for example, using a Synteni microarray (Palo Alto, Calif.) according to the manufacturer's instructions (and essentially as described by Schena et al., Proc. Natl. Acad. Sci. USA 93:10614-10619,1996 and Heller et al., Proc. Natl. Acad. Sci. USA 94:2150-2155,1997). Alternatively, polynucleotides may be amplified from cDNA prepared from cells expressing the proteins described herein. Such polynucleotides may be amplified via polymerase chain reaction (PCR). For this approach, sequence-specific primers may be designed based on the sequences provided herein, and may be purchased or synthesized.

[0233] An amplified portion of a polynucleotide of the present invention may be used to isolate a full length gene from a suitable library (e.g., an HSV cDNA library) using well known techniques. Within such techniques, a library (cDNA or genomic) is screened using one or more polynucleotide probes or primers suitable for amplification. Preferably, a library is size-selected to include larger molecules. Random primed libraries may also be preferred for identifying 5′ and upstream regions of genes. Genomic libraries are preferred for obtaining introns and extending 5′ sequences.

[0234] For hybridization techniques, a partial sequence may be labeled (e.g., by nick-translation or end-labeling with ³²P) using well known techniques. A bacterial or bacteriophage library is then generally screened by hybridizing filters containing denatured bacterial colonies (or lawns containing phage plaques) with the labeled probe (see Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratories, Cold Spring Harbor, N.Y., 1989). Hybridizing colonies or plaques are selected and expanded, and the DNA is isolated for further analysis. cDNA clones may be analyzed to determine the amount of additional sequence by, for example, PCR using a primer from the partial sequence and a primer from the vector. Restriction maps and partial sequences may be generated to identify one or more overlapping clones. The complete sequence may then be determined using standard techniques, which may involve generating a series of deletion clones. The resulting overlapping sequences can then assembled into a single contiguous sequence. A full length cDNA molecule can be generated by ligating suitable fragments, using well known techniques.

[0235] Alternatively, there are numerous amplification techniques for obtaining a full length coding sequence from a partial cDNA sequence. Within such techniques, amplification is generally performed via PCR. Any of a variety of commercially available kits may be used to perform the amplification step. Primers may be designed using, for example, software well known in the art. Primers are preferably 22-30 nucleotides in length, have a GC content of at least 50% and anneal to the target sequence at temperatures of about 68° C. to 72° C. The amplified region may be sequenced as described above, and overlapping sequences assembled into a contiguous sequence.

[0236] One such amplification technique is inverse PCR (see Triglia et al., Nucl. Acids Res. 16:8186, 1988), which uses restriction enzymes to generate a fragment in the known region of the gene. The fragment is then circularized by intramolecular ligation and used as a template for PCR with divergent primers derived from the known region. Within an alternative approach, sequences adjacent to a partial sequence may be retrieved by amplification with a primer to a linker sequence and a primer specific to a known region. The amplified sequences are typically subjected to a second round of amplification with the same linker primer and a second primer specific to the known region. A variation on this procedure, which employs two primers that initiate extension in opposite directions from the known sequence, is described in WO 96/38591. Another such technique is known as “rapid amplification of cDNA ends” or RACE. This technique involves the use of an internal primer and an external primer, which hybridizes to a polyA region or vector sequence, to identify sequences that are 5′ and 3′ of a known sequence. Additional techniques include capture PCR (Lagerstrom et al., PCR Methods Applic. 1:111-19, 1991) and walking PCR (Parker et al., Nucl. Acids. Res. 19:3055-60,1991). Other methods employing amplification may also be employed to obtain a full length cDNA sequence.

[0237] In certain instances, it is possible to obtain a full length cDNA sequence by analysis of sequences provided in an expressed sequence tag (EST) database, such as that available from GenBank. Searches for overlapping ESTs may generally be performed using well known programs (e.g., NCBI BLAST searches), and such ESTs may be used to generate a contiguous full length sequence. Full length DNA sequences may also be obtained by analysis of genomic fragments.

[0238] Polynucleotide Expression in Host Cells

[0239] In other embodiments of the invention, polynucleotide sequences or fragments thereof which encode polypeptides of the invention, or fusion proteins or functional equivalents thereof, may be used in recombinant DNA molecules to direct expression of a polypeptide in appropriate host cells. Due to the inherent degeneracy of the genetic code, other DNA sequences that encode substantially the same or a functionally equivalent amino acid sequence may be produced and these sequences may be used to clone and express a given polypeptide.

[0240] As will be understood by those of skill in the art, it may be advantageous in some instances to produce polypeptide-encoding nucleotide sequences possessing non-naturally occurring codons. For example, codons preferred by a particular prokaryotic or eukaryotic host can be selected to increase the rate of protein expression or to produce a recombinant RNA transcript having desirable properties, such as a half-life which is longer than that of a transcript generated from the naturally occurring sequence.

[0241] Moreover, the polynucleotide sequences of the present invention can be engineered using methods generally known in the art in order to alter polypeptide encoding sequences for a variety of reasons, including but not limited to, alterations which modify the cloning, processing, and/or expression of the gene product. For example, DNA shuffling by random fragmentation and PCR reassembly of gene fragments and synthetic oligonucleotides may be used to engineer the nucleotide sequences. In addition, site-directed mutagenesis may be used to insert new restriction sites, alter glycosylation patterns, change codon preference, produce splice variants, or introduce mutations, and so forth.

[0242] In another embodiment of the invention, natural, modified, or recombinant nucleic acid sequences may be ligated to a heterologous sequence to encode a fusion protein. For example, to screen peptide libraries for inhibitors of polypeptide activity, it may be useful to encode a chimeric protein that can be recognized by a commercially available antibody. A fusion protein may also be engineered to contain a cleavage site located between the polypeptide-encoding sequence and the heterologous protein sequence, so that the polypeptide may be cleaved and purified away from the heterologous moiety.

[0243] Sequences encoding a desired polypeptide may be synthesized, in whole or in part, using chemical methods well known in the art (see Caruthers, M. H. et al. (1980) Nucl. Acids Res. Symp. Ser. 215-223, Horn, T. et al. (1980) Nucl. Acids Res. Symp. Ser. 225-232). Alternatively, the protein itself may be produced using chemical methods to synthesize the amino acid sequence of a polypeptide, or a portion thereof. For example, peptide synthesis can be performed using various solid-phase techniques (Roberge, J. Y. et al. (1995) Science 269:202-204) and automated synthesis may be achieved, for example, using the ABI 431A Peptide Synthesizer (Perkin Elmer, Palo Alto, Calif.).

[0244] A newly synthesized peptide may be substantially purified by preparative high performance liquid chromatography (e.g., Creighton, T. (1983) Proteins, Structures and Molecular Principles, WH Freeman and Co., New York, N.Y.) or other comparable techniques available in the art. The composition of the synthetic peptides may be confirmed by amino acid analysis or sequencing (e.g., the Edman degradation procedure). Additionally, the amino acid sequence of a polypeptide, or any part thereof, may be altered during direct synthesis and/or combined using chemical methods with sequences from other proteins, or any part thereof, to produce a variant polypeptide.

[0245] In order to express a desired polypeptide, the nucleotide sequences encoding the polypeptide, or functional equivalents, may be inserted into appropriate expression vector, i.e., a vector which contains the necessary elements for the transcription and translation of the inserted coding sequence. Methods which are well known to those skilled in the art may be used to construct expression vectors containing sequences encoding a polypeptide of interest and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Such techniques are described in Sambrook, J. et al. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, Plainview, N.Y., and Ausubel, F. M. et al. (1989) Current Protocols in Molecular Biology, John Wiley & Sons, New York. N.Y.

[0246] A variety of expression vector/host systems may be utilized to contain and express polynucleotide sequences. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with virus expression vectors (e.g., baculovirus); plant cell systems transformed with virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems.

[0247] The “control elements” or “regulatory sequences” present in an expression vector are those non-translated regions of the vector—enhancers, promoters, 5′ and 3′ untranslated regions—which interact with host cellular proteins to carry out transcription and translation. Such elements may vary in their strength and specificity. Depending on the vector system and host utilized, any number of suitable transcription and translation elements, including constitutive and inducible promoters, may be used. For example, when cloning in bacterial systems, inducible promoters such as the hybrid lacZ promoter of the PBLUESCRIPT phagemid (Stratagene, La Jolla, Calif.) or PSPORT1 plasmid (Gibco BRL, Gaithersburg, Md.) and the like may be used. In mammalian cell systems, promoters from mammalian genes or from mammalian viruses are generally preferred. If it is necessary to generate a cell line that contains multiple copies of the sequence encoding a polypeptide, vectors based on SV40 or EBV may be advantageously used with an appropriate selectable marker.

[0248] In bacterial systems, a number of expression vectors may be selected depending upon the use intended for the expressed polypeptide. For example, when large quantities are needed, for example for the induction of antibodies, vectors which direct high level expression of fusion proteins that are readily purified may be used. Such vectors include, but are not limited to, the multifunctional E. coli cloning and expression vectors such as BLUESCRIPT (Stratagene), in which the sequence encoding the polypeptide of interest may be ligated into the vector in frame with sequences for the amino-terminal Met and the subsequent 7 residues of .beta.-galactosidase so that a hybrid protein is produced; pIN vectors (Van Heeke, G. and S. M. Schuster (1989) J. Biol. Chem. 264:5503-5509); and the like. pGEX Vectors (Promega, Madison, Wis.) may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins made in such systems may be designed to include heparin, thrombin, or factor XA protease cleavage sites so that the cloned polypeptide of interest can be released from the GST moiety at will.

[0249] In the yeast, Saccharomyces cerevisiae, a number of vectors containing constitutive or inducible promoters such as alpha factor, alcohol oxidase, and PGH may be used. For reviews, see Ausubel et al. (supra) and Grant et al. (1987) Methods Enzymol. 153:516-544.

[0250] In cases where plant expression vectors are used, the expression of sequences encoding polypeptides may be driven by any of a number of promoters. For example, viral promoters such as the 35S and 19S promoters of CaMV may be used alone or in combination with the omega leader sequence from TMV (Takamatsu, N. (1987) EMBO J. 6:307-311. Alternatively, plant promoters such as the small subunit of RUBISCO or heat shock promoters may be used (Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680; Broglie, R. et al. (1984) Science 224:838-843; and Winter, J. et al. (1991) Results Probl. Cell Differ. 17:85-105). These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. Such techniques are described in a number of generally available reviews (see, for example, Hobbs, S. or Murry, L. E. in McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York, N.Y.; pp. 191-196).

[0251] An insect system may also be used to express a polypeptide of interest. For example, in one such system, Autographa californica nuclear polyhedrosis virus (ACNPV) is used as a vector to express foreign genes in Spodoptera frugiperda cells or in Trichoplusia larvae. The sequences encoding the polypeptide may be cloned into a non-essential region of the virus, such as the polyhedrin gene, and placed under control of the polyhedrin promoter. Successful insertion of the polypeptide-encoding sequence will render the polyhedrin gene inactive and produce recombinant virus lacking coat protein. The recombinant viruses may then be used to infect, for example, S. frugiperda cells or Trichoplusia larvae in which the polypeptide of interest may be expressed (Engelhard, E. K. et al. (1994) Proc. Natl. Acad. Sci. 91:3224-3227).

[0252] In mammalian host cells, a number of viral-based expression systems are generally available. For example, in cases where an adenovirus is used as an expression vector, sequences encoding a polypeptide of interest may be ligated into an adenovirus transcription/translation complex consisting of the late promoter and tripartite leader sequence. Insertion in a non-essential E1 or E3 region of the viral genome may be used to obtain a viable virus which is capable of expressing the polypeptide in infected host cells (Logan, J. and Shenk, T. (1984) Proc. Natl. Acad. Sci. 81:3655-3659). In addition, transcription enhancers, such as the Rous sarcoma virus (RSV) enhancer, may be used to increase expression in mammalian host cells.

[0253] Specific initiation signals may also be used to achieve more efficient translation of sequences encoding a polypeptide of interest. Such signals include the ATG initiation codon and adjacent sequences. In cases where sequences encoding the polypeptide, its initiation codon, and upstream sequences are inserted into the appropriate expression vector, no additional transcriptional or translational control signals may be needed. However, in cases where only coding sequence, or a portion thereof, is inserted, exogenous translational control signals including the ATG initiation codon should be provided. Furthermore, the initiation codon should be in the correct reading frame to ensure translation of the entire insert. Exogenous translational elements and initiation codons may be of various origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of enhancers which are appropriate for the particular cell system which is used, such as those described in the literature (Scharf, D. et al. (1994) Results Probl. Cell Differ. 20:125-162).

[0254] In addition, a host cell strain may be chosen for its ability to modulate the expression of the inserted sequences or to process the expressed protein in the desired fashion. Such modifications of the polypeptide include, but are not limited to, acetylation, carboxylation. glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing which cleaves a “prepro” form of the protein may also be used to facilitate correct insertion, folding and/or function. Different host cells such as CHO, HeLa, MDCK, HEK293, and W138, which have specific cellular machinery and characteristic mechanisms for such post-translational activities, may be chosen to ensure the correct modification and processing of the foreign protein.

[0255] For long-term, high-yield production of recombinant proteins, stable expression is generally preferred. For example, cell lines which stably express a polynucleotide of interest may be transformed using expression vectors which may contain viral origins of replication and/or endogenous expression elements and a selectable marker gene on the same or on a separate vector. Following the introduction of the vector, cells may be allowed to grow for 1-2 days in an enriched media before they are switched to selective media. The purpose of the selectable marker is to confer resistance to selection, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clones of stably transformed cells may be proliferated using tissue culture techniques appropriate to the cell type.

[0256] Any number of selection systems may be used to recover transformed cell lines. These include, but are not limited to, the herpes simplex virus thymidine kinase (Wigler, M. et al. (1977) Cell 11:223-32) and adenine phosphoribosyltransferase (Lowy, I. et al. (1990) Cell 22:817-23) genes which can be employed in tk.sup.- or aprt.sup.- cells, respectively. Also, antimetabolite, antibiotic or herbicide resistance can be used as the basis for selection; for example, dhfr which confers resistance to methotrexate (Wigler, M. et al. (1980) Proc. Natl. Acad. Sci. 77:3567-70); npt, which confers resistance to the aminoglycosides, neomycin and G-418 (Colbere-Garapin, F. et al (1981) J. Mol. Biol. 150:1-14); and als or pat, which confer resistance to chlorsulfuron and phosphinotricin acetyltransferase, respectively (Murry, supra). Additional selectable genes have been described, for example, trpB, which allows cells to utilize indole in place of tryptophan, or hisD, which allows cells to utilize histinol in place of histidine (Hartman, S. C. and R. C. Mulligan (1988) Proc. Natl. Acad. Sci. 85:8047-51). Recently, the use of visible markers has gained popularity with such markers as anthocyanins, beta-glucuronidase and its substrate GUS, and luciferase and its substrate luciferin, being widely used not only to identify transformants, but also to quantify the amount of transient or stable protein expression attributable to a specific vector system (Rhodes, C. A. et al. (1995) Methods Mol. Biol. 55:121-131).

[0257] Although the presence/absence of marker gene expression suggests that the gene of interest is also present, its presence and expression may need to be confirmed. For example, if the sequence encoding a polypeptide is inserted within a marker gene sequence, recombinant cells containing sequences can be identified by the absence of marker gene function. Alternatively, a marker gene can be placed in tandem with a polypeptide-encoding sequence under the control of a single promoter. Expression of the marker gene in response to induction or selection usually indicates expression of the tandem gene as well.

[0258] Alternatively, host cells which contain and express a desired polynucleotide sequence may be identified by a variety of procedures known to those of skill in the art. These procedures include, but are not limited to, DNA-DNA or DNA-RNA hybridizations and protein bioassay or immunoassay techniques which include membrane, solution, or chip based technologies for the detection and/or quantification of nucleic acid or protein.

[0259] A variety of protocols for detecting and measuring the expression of polynucleotide-encoded products, using either polyclonal or monoclonal antibodies specific for the product are known in the art. Examples include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and fluorescence activated cell sorting (FACS). A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes on a given polypeptide may be preferred for some applications, but a competitive binding assay may also be employed. These and other assays are described, among other places, in Hampton, R. et al. (1990; Serological Methods, a Laboratory Manual, APS Press, St Paul. Minn.) and Maddox, D. E. et al. (1983; J. Exp. Med. 158:1211-1216).

[0260] A wide variety of labels and conjugation techniques are known by those skilled in the art and may be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting sequences related to polynucleotides include oligolabeling, nick translation, end-labeling or PCR amplification using a labeled nucleotide. Alternatively, the sequences, or any portions thereof may be cloned into a vector for the production of an mRNA probe. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by addition of an appropriate RNA polymerase such as T7, T3, or SP6 and labeled nucleotides. These procedures may be conducted using a variety of commercially available kits. Suitable reporter molecules or labels, which may be used include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents as well as substrates, cofactors, inhibitors, magnetic particles, and the like.

[0261] Host cells transformed with a polynucleotide sequence of interest may be cultured under conditions suitable for the expression and recovery of the protein from cell culture. The protein produced by a recombinant cell may be secreted or contained intracellularly depending on the sequence and/or the vector used. As will be understood by those of skill in the art, expression vectors containing polynucleotides of the invention may be designed to contain signal sequences which direct secretion of the encoded polypeptide through a prokaryotic or eukaryotic cell membrane. Other recombinant constructions may be used to join sequences encoding a polypeptide of interest to nucleotide sequence encoding a polypeptide domain which will facilitate purification of soluble proteins. Such purification facilitating domains include, but are not limited to, metal chelating peptides such as histidine-tryptophan modules that allow purification on immobilized metals, protein A domains that allow purification on immobilized immunoglobulin, and the domain utilized in the FLAGS extension/affinity purification system (Immunex Corp., Seattle, Wash.). The inclusion of cleavable linker sequences such as those specific for Factor XA or enterokinase (Invitrogen. San Diego, Calif.) between the purification domain and the encoded polypeptide may be used to facilitate purification. One such expression vector provides for expression of a fusion protein containing a polypeptide of interest and a nucleic acid encoding 6 histidine residues preceding a thioredoxin or an enterokinase cleavage site. The histidine residues facilitate purification on IMIAC (immobilized metal ion affinity chromatography) as described in Porath, J. et al. (1992, Prot. Exp. Purif. 3:263-281) while the enterokinase cleavage site provides a means for purifying the desired polypeptide from the fusion protein. A discussion of vectors which contain fusion proteins is provided in Kroll, D. J. et al. (1993; DNA Cell Biol. 12:441-453).

[0262] In addition to recombinant production methods, polypeptides of the invention, and fragments thereof, may be produced by direct peptide synthesis using solid-phase techniques (Merrifield J. (1963) J. Am. Chem. Soc. 85:2149-2154). Protein synthesis may be performed using manual techniques or by automation. Automated synthesis may be achieved, for example, using Applied Biosystems 431A Peptide Synthesizer (Perkin Elmer). Alternatively, various fragments may be chemically synthesized separately and combined using chemical methods to produce the full length molecule.

[0263] Site-Specific Mutagenesis

[0264] Site-specific mutagenesis is a technique useful in the preparation of individual peptides, or biologically functional equivalent polypeptides, through specific mutagenesis of the underlying polynucleotides that encode them. The technique, well-known to those of skill in the art, further provides a ready ability to prepare and test sequence variants, for example, incorporating one or more of the foregoing considerations, by introducing one or more nucleotide sequence changes into the DNA. Site-specific mutagenesis allows the production of mutants through the use of specific oligonucleotide sequences which encode the DNA sequence of the desired mutation, as well as a sufficient number of adjacent nucleotides, to provide a primer sequence of sufficient size and sequence complexity to form a stable duplex on both sides of the deletion junction being traversed. Mutations may be employed in a selected polynucleotide sequence to improve, alter, decrease, modify, or otherwise change the properties of the polynucleotide itself, and/or alter the properties, activity, composition, stability, or primary sequence of the encoded polypeptide.

[0265] In certain embodiments of the present invention, the inventors contemplate the mutagenesis of the disclosed polynucleotide sequences to alter one or more properties of the encoded polypeptide, such as the antigenicity of a polypeptide vaccine. The techniques of site-specific mutagenesis are well-known in the art, and are widely used to create variants of both polypeptides and polynucleotides. For example, site-specific mutagenesis is often used to alter a specific portion of a DNA molecule. In such embodiments, a primer comprising typically about 14 to about 25 nucleotides or so in length is employed, with about 5 to about 10 residues on both sides of the junction of the sequence being altered.

[0266] As will be appreciated by those of skill in the art, site-specific mutagenesis techniques have often employed a phage vector that exists in both a single stranded and double stranded form. Typical vectors useful in site-directed mutagenesis include vectors such as the M13 phage. These phage are readily commercially-available and their use is generally well-known to those skilled in the art. Double-stranded plasmids are also routinely employed in site directed mutagenesis that eliminates the step of transferring the gene of interest from a plasmid to a phage.

[0267] In general, site-directed mutagenesis in accordance herewith is performed by first obtaining a single-stranded vector or melting apart of two strands of a double-stranded vector that includes within its sequence a DNA sequence that encodes the desired peptide. An oligonucleotide primer bearing the desired mutated sequence is prepared, generally synthetically. This primer is then annealed with the single-stranded vector, and subjected to DNA polymerizing enzymes such as E. coli polymerase I Klenow fragment, in order to complete the synthesis of the mutation-bearing strand. Thus, a heteroduplex is formed wherein one strand encodes the original non-mutated sequence and the second strand bears the desired mutation. This heteroduplex vector is then used to transform appropriate cells, such as E. coli cells, and clones are selected which include recombinant vectors bearing the mutated sequence arrangement.

[0268] The preparation of sequence variants of the selected peptide-encoding DNA segments using site-directed mutagenesis provides a means of producing potentially useful species and is not meant to be limiting as there are other ways in which sequence variants of peptides and the DNA sequences encoding them may be obtained. For example, recombinant vectors encoding the desired peptide sequence may be treated with mutagenic agents, such as hydroxylamine, to obtain sequence variants. Specific details regarding these methods and protocols are found in the teachings of Maloy et al., 1994; Segal, 1976; Prokop and Bajpai, 1991; Kuby, 1994; and Maniatis et al., 1982, each incorporated herein by reference, for that purpose.

[0269] As used herein, the term “oligonucleotide directed mutagenesis procedure” refers to template-dependent processes and vector-mediated propagation which result in an increase in the concentration of a specific nucleic acid molecule relative to its initial concentration, or in an increase in the concentration of a detectable signal, such as amplification. As used herein, the term “oligonucleotide directed mutagenesis procedure” is intended to refer to a process that involves the template-dependent extension of a primer molecule. The term template dependent process refers to nucleic acid synthesis of an RNA or a DNA molecule wherein the sequence of the newly synthesized strand of nucleic acid is dictated by the well-known rules of complementary base pairing (see, for example, Watson, 1987). Typically, vector mediated methodologies involve the introduction of the nucleic acid fragment into a DNA or RNA vector, the clonal amplification of the vector, and the recovery of the amplified nucleic acid fragment. Examples of such methodologies are provided by U.S. Pat. No. 4,237,224, specifically incorporated herein by reference in its entirety.

[0270] Polynucleotide Amplification Techniques

[0271] A number of template dependent processes are available to amplify the target sequences of interest present in a sample. One of the best known amplification methods is the polymerase chain reaction (PCR™) which is described in detail in U.S. Pat. Nos. 4,683,195, 4,683,202 and 4,800,159, each of which is incorporated herein by reference in its entirety. Briefly, in PCR™, two primer sequences are prepared which are complementary to regions on opposite complementary strands of the target sequence. An excess of deoxynucleoside triphosphates is added to a reaction mixture along with a DNA polymerase (e.g., Taq polymerase). If the target sequence is present in a sample, the primers will bind to the target and the polymerase will cause the primers to be extended along the target sequence by adding on nucleotides. By raising and lowering the temperature of the reaction mixture, the extended primers will dissociate from the target to form reaction products, excess primers will bind to the target and to the reaction product and the process is repeated. Preferably reverse transcription and PCR™ amplification procedure may be performed in order to quantify the amount of mRNA amplified. Polymerase chain reaction methodologies are well known in the art.

[0272] Another method for amplification is the ligase chain reaction (referred to as LCR), disclosed in Eur. Pat. Appl. Publ. No. 320,308 (specifically incorporated herein by reference in its entirety). In LCR, two complementary probe pairs are prepared, and in the presence of the target sequence, each pair will bind to opposite complementary strands of the target such that they abut. In the presence of a ligase, the two probe pairs will link to form a single unit. By temperature cycling, as in PCR™, bound ligated units dissociate from the target and then serve as “target sequences” for ligation of excess probe pairs. U.S. Pat. No. 4,883,750, incorporated herein by reference in its entirety, describes an alternative method of amplification similar to LCR for binding probe pairs to a target sequence.

[0273] Qbeta Replicase, described in PCT Intl. Pat. Appl. Publ. No. PCT/US87/00880, incorporated herein by reference in its entirety, may also be used as still another amplification method in the present invention. In this method, a replicative sequence of RNA that has a region complementary to that of a target is added to a sample in the presence of an RNA polymerase. The polymerase will copy the replicative sequence that can then be detected.

[0274] An isothermal amplification method, in which restriction endonucleases and ligases are used to achieve the amplification of target molecules that contain nucleotide 5′-[α-thio]triphosphates in one strand of a restriction site (Walker et al., 1992, incorporated herein by reference in its entirety), may also be useful in the amplification of nucleic acids in the present invention.

[0275] Strand Displacement Amplification (SDA) is another method of carrying out isothermal amplification of nucleic acids which involves multiple rounds of strand displacement and synthesis, i.e. nick translation. A similar method, called Repair Chain Reaction (RCR) is another method of amplification which may be useful in the present invention and is involves annealing several probes throughout a region targeted for amplification, followed by a repair reaction in which only two of the four bases are present. The other two bases can be added as biotinylated derivatives for easy detection. A similar approach is used in SDA.

[0276] Sequences can also be detected using a cyclic probe reaction (CPR). In CPR, a probe having a 3′ and 5′ sequences of non-target DNA and an internal or “middle” sequence of the target protein specific RNA is hybridized to DNA which is present in a sample. Upon hybridization, the reaction is treated with RNaseH, and the products of the probe are identified as distinctive products by generating a signal that is released after digestion. The original template is annealed to another cycling probe and the reaction is repeated. Thus, CPR involves amplifying a signal generated by hybridization of a probe to a target gene specific expressed nucleic acid.

[0277] Still other amplification methods described in Great Britain Pat. Appl. No. 2 202 328, and in PCT Intl. Pat. Appl. Publ. No. PCT/US89/01025, each of which is incorporated herein by reference in its entirety, may be used in accordance with the present invention. In the former application, “modified” primers are used in a PCR-like, template and enzyme dependent synthesis. The primers may be modified by labeling with a capture moiety (e.g., biotin) and/or a detector moiety (e.g., enzyme). In the latter application, an excess of labeled probes is added to a sample. In the presence of the target sequence, the probe binds and is cleaved catalytically. After cleavage, the target sequence is released intact to be bound by excess probe. Cleavage of the labeled probe signals the presence of the target sequence.

[0278] Other nucleic acid amplification procedures include transcription-based amplification systems (TAS) (Kwoh et al., 1989; PCT Intl. Pat. Appl. Publ. No. WO 88/10315, incorporated herein by reference in its entirety), including nucleic acid sequence based amplification (NASBA) and 3SR. In NASBA, the nucleic acids can be prepared for amplification by standard phenol/chloroform extraction, heat denaturation of a sample, treatment with lysis buffer and minispin columns for isolation of DNA and RNA or guanidinium chloride extraction of RNA. These amplification techniques involve annealing a primer that has sequences specific to the target sequence. Following polymerization, DNA/RNA hybrids are digested with RNase H while double stranded DNA molecules are heat-denatured again. In either case the single stranded DNA is made fully double stranded by addition of second target-specific primer, followed by polymerization. The double stranded DNA molecules are then multiply transcribed by a polymerase such as T7 or SP6. In an isothermal cyclic reaction, the RNAs are reverse transcribed into DNA, and transcribed once again with a polymerase such as T7 or SP6. The resulting products, whether truncated or complete, indicate target-specific sequences.

[0279] Eur. Pat. Appl. Publ. No. 329,822, incorporated herein by reference in its entirety, disclose a nucleic acid amplification process involving cyclically synthesizing single-stranded RNA (“ssRNA”), ssDNA, and double-stranded DNA (dsDNA), which may be used in accordance with the present invention. The ssRNA is a first template for a first primer oligonucleotide, which is elongated by reverse transcriptase (RNA-dependent DNA polymerase). The RNA is then removed from resulting DNA: RNA duplex by the action of ribonuclease H (RNase H, an RNase specific for RNA in a duplex with either DNA or RNA). The resultant ssDNA is a second template for a second primer, which also includes the sequences of an RNA polymerase promoter (exemplified by T7 RNA polymerase) 5′ to its homology to its template. This primer is then extended by DNA polymerase (exemplified by the large “Klenow” fragment of E. coli DNA polymerase 1), resulting as a double-stranded DNA (“dsDNA”) molecule, having a sequence identical to that of the original RNA between the primers and having additionally, at one end, a promoter sequence. This promoter sequence can be used by the appropriate RNA polymerase to make many RNA copies of the DNA. These copies can then re-enter the cycle leading to very swift amplification. With proper choice of enzymes, this amplification can be done isothermally without addition of enzymes at each cycle. Because of the cyclical nature of this process, the starting sequence can be chosen to be in the form of either DNA or RNA.

[0280] PCT Intl. Pat. Appl. Publ. No. WO 89/06700, incorporated herein by reference in its entirety, disclose a nucleic acid sequence amplification scheme based on the hybridization of a promoter/primer sequence to a target single-stranded DNA (“ssDNA”) followed by transcription of many RNA copies of the sequence. This scheme is not cyclic; i.e., new templates are not produced from the resultant RNA transcripts. Other amplification methods include “RACE” (Frohman, 1990), and “one-sided PCR” (Ohara, 1989) which are well-known to those of skill in the art.

[0281] Methods based on ligation of two (or more) oligonucleotides in the presence of nucleic acid having the sequence of the resulting “di-oligonucleotide”, thereby amplifying the di-oligonucleotide (Wu and Dean, 1996, incorporated herein by reference in its entirety), may also be used in the amplification of DNA sequences of the present invention.

[0282] Biological Functional Equivalents

[0283] Modification and changes may be made in the structure of the polynucleotides and polypeptides of the present invention and still obtain a functional molecule that encodes a polypeptide with desirable characteristics. As mentioned above, it is often desirable to introduce one or more mutations into a specific polynucleotide sequence. In certain circumstances, the resulting encoded polypeptide sequence is altered by this mutation, or in other cases, the sequence of the polypeptide is unchanged by one or more mutations in the encoding polynucleotide.

[0284] When it is desirable to alter the amino acid sequence of a polypeptide to create an equivalent, or even an improved, second-generation molecule, the amino acid changes may be achieved by changing one or more of the codons of the encoding DNA sequence, according to Table 1.

[0285] For example, certain amino acids may be substituted for other amino acids in a protein structure without appreciable loss of interactive binding capacity with structures such as, for example, antigen-binding regions of antibodies or binding sites on substrate molecules. Since it is the interactive capacity and nature of a protein that defines that protein's biological functional activity, certain amino acid sequence substitutions can be made in a protein sequence, and, of course, its underlying DNA coding sequence, and nevertheless obtain a protein with like properties. It is thus contemplated by the inventors that various changes may be made in the peptide sequences of the disclosed compositions, or corresponding DNA sequences which encode said peptides without appreciable loss of their biological utility or activity. TABLE 1 Amino Acids Codons Alanine Ala A GCA GCC GCG GCU Cysteine Cys C UGC UGU Aspartic acid Asp D GAC GAU GLutamic acid Glu E GAA GAG Phenylalanine Phe F UUC UUU Glycine Gly G GGA GGC GGG GGU Histidine His H CAC CAU Isoleucine Ile I AUA AUC AUU Lysine Lys K AAA AAG Leucine Leu L UUA UUG CUA CUC CUG CUU Methionine Met M AUG Asparagine Asn N AAC AAU Proline Pro P CCA CCC CCG CCU Glutamine Gln Q CAA CAG Arginine Arg R AGA AGG CGA CGC CGG CGU Serine Ser S AGC AGU UCA UCC UCG UCU Threonine Thr T ACA ACC ACG ACU Valine Val V GUA GUC GUG GUU Tryptophan Trp W UGG Tyrosine Tyr Y UAC UAU

[0286] In making such changes, the hydropathic index of amino acids may be considered. The importance of the hydropathic amino acid index in conferring interactive biologic function on a protein is generally understood in the art (Kyte and Doolittle, 1982, incorporated herein by reference). It is accepted that the relative hydropathic character of the amino acid contributes to the secondary structure of the resultant protein, which in turn defines the interaction of the protein with other molecules, for example, enzymes, substrates, receptors, DNA, antibodies, antigens, and the like. Each amino acid has been assigned a hydropathic index on the basis of its hydrophobicity and charge characteristics (Kyte and Doolittle, 1982). These values are: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (−0.4); threonine (−0.7); serine (−0.8); tryptophan (−0.9); tyrosine (−1.3); proline (−1.6); histidine (−3.2); glutamate (−3.5); glutamine (−3.5); aspartate (−3.5); asparagine (−3.5); lysine (−3.9); and arginine (−4.5).

[0287] It is known in the art that certain amino acids may be substituted by other amino acids having a similar hydropathic index or score and still result in a protein with similar biological activity, i.e., still obtain a biological functionally equivalent protein. In making such changes, the substitution of amino acids whose hydropathic indices are within ±2 is preferred, those within ±1 are particularly preferred, and those within ±0.5 are even more particularly preferred. It is also understood in the art that the substitution of like amino acids can be made effectively on the basis of hydrophilicity. U.S. Pat. No. 4,554,101 (specifically incorporated herein by reference in its entirety), states that the greatest local average hydrophilicity of a protein, as governed by the hydrophilicity of its adjacent amino acids, correlates with a biological property of the protein.

[0288] As detailed in U.S. Pat. No. 4,554,101, the following hydrophilicity values have been assigned to amino acid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0±1); glutamate (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (−0.4); proline (−0.5 +1); alanine (−0.5); histidine (−0.5); cysteine (−1.0); methionine (−1.3); valine (−1.5); leucine (−1.8); isoleucine (−1.8); tyrosine (−2.3); phenylalanine (−2.5); tryptophan (−3.4). It is understood that an amino acid can be substituted for another having a similar hydrophilicity value and still obtain a biologically equivalent, and in particular, an immunologically equivalent protein. In such changes, the substitution of amino acids whose hydrophilicity values are within ±2 is preferred, those within ±1 are particularly preferred, and those within ±0.5 are even more particularly preferred.

[0289] As outlined above, amino acid substitutions are generally therefore based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity, hydrophilicity, charge, size, and the like. Exemplary substitutions that take various of the foregoing characteristics into consideration are well known to those of skill in the art and include: arginine and lysine; glutamate and aspartate; serine and threonine; glutamine and asparagine; and valine, leucine and isoleucine.

[0290] In addition, any polynucleotide may be further modified to increase stability in vivo. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5′ and/or 3′ ends; the use of phosphorothioate or 2′ O-methyl rather than phosphodiesterase linkages in the backbone; and/or the inclusion of nontraditional bases such as inosine, queosine and wybutosine, as well as acetyl- methyl-, thio- and other modified forms of adenine, cytidine, guanine, thymine and uridine.

[0291] In vivo Polynucleotide Delivery Techniques

[0292] In additional embodiments, genetic constructs comprising one or more of the polynucleotides of the invention are introduced into cells in vivo. This may be achieved using any of a variety or well known approaches, several of which are outlined below for the purpose of illustration.

[0293] 1. Adenovirus

[0294] One of the preferred methods for in vivo delivery of one or more nucleic acid sequences involves the use of an adenovirus expression vector. “Adenovirus expression vector” is meant to include those constructs containing adenovirus sequences sufficient to (a) support packaging of the construct and (b) to express a polynucleotide that has been cloned therein in a sense or antisense orientation. Of course, in the context of an antisense construct, expression does not require that the gene product be synthesized.

[0295] The expression vector comprises a genetically engineered form of an adenovirus. Knowledge of the genetic organization of adenovirus, a 36 kb, linear, double-stranded DNA virus, allows substitution of large pieces of adenoviral DNA with foreign sequences up to 7 kb (Grunhaus and Horwitz, 1992). In contrast to retrovirus, the adenoviral infection of host cells does not result in chromosomal integration because adenoviral DNA can replicate in an episomal manner without potential genotoxicity. Also, adenoviruses are structurally stable, and no genome rearrangement has been detected after extensive amplification. Adenovirus can infect virtually all epithelial cells regardless of their cell cycle stage. So far, adenoviral infection appears to be linked only to mild disease such as acute respiratory disease in humans.

[0296] Adenovirus is particularly suitable for use as a gene transfer vector because of its mid-sized genome, ease of manipulation, high titer, wide target-cell range and high infectivity. Both ends of the viral genome contain 100-200 base pair inverted repeats (ITRs), which are cis elements necessary for viral DNA replication and packaging. The early (E) and late (L) regions of the genome contain different transcription units that are divided by the onset of viral DNA replication. The E1 region (E1A and E1B) encodes proteins responsible for the regulation of transcription of the viral genome and a few cellular genes. The expression of the E2 region (E2A and E2B) results in the synthesis of the proteins for viral DNA replication. These proteins are involved in DNA replication, late gene expression and host cell shut-off (Renan, 1990). The products of the late genes, including the majority of the viral capsid proteins, are expressed only after significant processing of a single primary transcript issued by the major late promoter (MLP). The MLP, (located at 16.8 m.u.) is particularly efficient during the late phase of infection, and all the mRNA's issued from this promoter possess a 5′-tripartite leader (TPL) sequence which makes them preferred mRNA's for translation.

[0297] In a current system, recombinant adenovirus is generated from homologous recombination between shuttle vector and provirus vector. Due to the possible recombination between two proviral vectors, wild-type adenovirus may be generated from this process. Therefore, it is critical to isolate a single clone of virus from an individual plaque and examine its genomic structure.

[0298] Generation and propagation of the current adenovirus vectors, which are replication deficient, depend on a unique helper cell line, designated 293, which was transformed from human embryonic kidney cells by Ad5 DNA fragments and constitutively expresses E1 proteins (Graham et al., 1977). Since the E3 region is dispensable from the adenovirus genome (Jones and Shenk, 1978), the current adenovirus vectors, with the help of 293 cells, carry foreign DNA in either the E1, the D3 or both regions (Graham and Prevec, 1991). In nature, adenovirus can package approximately 105% of the wild-type genome (Ghosh-Choudhury et al., 1987), providing capacity for about 2 extra kB of DNA. Combined with the approximately 5.5 kB of DNA that is replaceable in the E1 and E3 regions, the maximum capacity of the current adenovirus vector is under 7.5 kB, or about 15% of the total length of the vector. More than 80% of the adenovirus viral genome remains in the vector backbone and is the source of vector-borne cytotoxicity. Also, the replication deficiency of the E1-deleted virus is incomplete. For example, leakage of viral gene expression has been observed with the currently available vectors at high multiplicities of infection (MOI) (Mulligan, 1993).

[0299] Helper cell lines may be derived from human cells such as human embryonic kidney cells, muscle cells, hematopoietic cells or other human embryonic mesenchymal or epithelial cells. Alternatively, the helper cells may be derived from the cells of other mammalian species that are permissive for human adenovirus. Such cells include, e.g., Vero cells or other monkey embryonic mesenchymal or epithelial cells. As stated above, the currently preferred helper cell line is 293.

[0300] Recently, Racher et al. (1995) disclosed improved methods for culturing 293 cells and propagating adenovirus. In one format, natural cell aggregates are grown by inoculating individual cells into 1 liter siliconized spinner flasks (Techne, Cambridge, UK) containing 100-200 ml of medium. Following stirring at 40 rpm, the cell viability is estimated with trypan blue. In another format, Fibra-Cel microcarriers (Bibby Sterlin, Stone, UK) (5 g/l) is employed as follows. A cell inoculum, resuspended in 5 ml of medium, is added to the carrier (50 ml) in a 250 ml Erlenmeyer flask and left stationary, with occasional agitation, for 1 to 4 h. The medium is then replaced with 50 ml of fresh medium and shaking initiated. For virus production, cells are allowed to grow to about 80% confluence, after which time the medium is replaced (to 25% of the final volume) and adenovirus added at an MOI of 0.05. Cultures are left stationary overnight, following which the volume is increased to 100% and shaking commenced for another 72 h.

[0301] Other than the requirement that the adenovirus vector be replication defective, or at least conditionally defective, the nature of the adenovirus vector is not believed to be crucial to the successful practice of the invention. The adenovirus may be of any of the 42 different known serotypes or subgroups A-F. Adenovirus type 5 of subgroup C is the preferred starting material in order to obtain a conditional replication-defective adenovirus vector for use in the present invention, since Adenovirus type 5 is a human adenovirus about which a great deal of biochemical and genetic information is known, and it has historically been used for most constructions employing adenovirus as a vector.

[0302] As stated above, the typical vector according to the present invention is replication defective and will not have an adenovirus El region. Thus, it will be most convenient to introduce the polynucleotide encoding the gene of interest at the position from which the E1-coding sequences have been removed. However, the position of insertion of the construct within the adenovirus sequences is not critical to the invention. The polynucleotide encoding the gene of interest may also be inserted in lieu of the deleted E3 region in E3 replacement vectors as described by Karlsson et al. (1986) or in the E4 region where a helper cell line or helper virus complements the E4 defect.

[0303] Adenovirus is easy to grow and manipulate and exhibits broad host range in vitro and in vivo. This group of viruses can be obtained in high titers, e.g., 10⁹-10¹¹ plaque-forming units per ml, and they are highly infective. The life cycle of adenovirus does not require integration into the host cell genome. The foreign genes delivered by adenovirus vectors are episomal and, therefore, have low genotoxicity to host cells. No side effects have been reported in studies of vaccination with wild-type adenovirus (Couch et al., 1963; Top et al., 1971), demonstrating their safety and therapeutic potential as in vivo gene transfer vectors.

[0304] Adenovirus vectors have been used in eukaryotic gene expression (Levrero et al., 1991; Gomez-Foix et al., 1992) and vaccine development (Grunhaus and Horwitz, 1992; Graham and Prevec, 1992). Recently, animal studies suggested that recombinant adenovirus could be used for gene therapy (Strafford-Perricaudet and Perricaudet, 1991; Stratford-Perricaudet et al., 1990; Rich et al., 1993). Studies in administering recombinant adenovirus to different tissues include trachea instillation (Rosenfeld et al., 1991; Rosenfeld et al., 1992), muscle injection (Ragot et al., 1993), peripheral intravenous injections (Herz and Gerard, 1993) and stereotactic inoculation into the brain (Le Gal La Salle et al., 1993).

[0305] 2. Retroviruses

[0306] The retroviruses are a group of single-stranded RNA viruses characterized by an ability to convert their RNA to double-stranded DNA in infected cells by a process of reverse-transcription (Coffin, 1990). The resulting DNA then stably integrates into cellular chromosomes as a provirus and directs synthesis of viral proteins. The integration results in the retention of the viral gene sequences in the recipient cell and its descendants. The retroviral genome contains three genes, gag, pol, and env that code for capsid proteins, polymerase enzyme, and envelope components, respectively. A sequence found upstream from the gag gene contains a signal for packaging of the genome into virions. Two long terminal repeat (LTR) sequences are present at the 5′ and 3′ ends of the viral genome. These contain strong promoter and enhancer sequences and are also required for integration in the host cell genome (Coffin, 1990).

[0307] In order to construct a retroviral vector, a nucleic acid encoding one or more oligonucleotide or polynucleotide sequences of interest is inserted into the viral genome in the place of certain viral sequences to produce a virus that is replication-defective. In order to produce virions, a packaging cell line containing the gag, pol, and env genes but without the LTR and packaging components is constructed (Mann et al., 1983). When a recombinant plasmid containing a cDNA, together with the retroviral LTR and packaging sequences is introduced into this cell line (by calcium phosphate precipitation for example), the packaging sequence allows the RNA transcript of the recombinant plasmid to be packaged into viral particles, which are then secreted into the culture media (Nicolas and Rubenstein, 1988; Temin, 1986; Mann et al., 1983). The media containing the recombinant retroviruses is then collected, optionally concentrated, and used for gene transfer. Retroviral vectors are able to infect a broad variety of cell types. However, integration and stable expression require the division of host cells (Paskind et al., 1975).

[0308] A novel approach designed to allow specific targeting of retrovirus vectors was recently developed based on the chemical modification of a retrovirus by the chemical addition of lactose residues to the viral envelope. This modification could permit the specific infection of hepatocytes via sialoglycoprotein receptors.

[0309] A different approach to targeting of recombinant retroviruses was designed in which biotinylated antibodies against a retroviral envelope protein and against a specific cell receptor were used. The antibodies were coupled via the biotin components by using streptavidin (Roux et al., 1989). Using antibodies against major histocompatibility complex class I and class II antigens, they demonstrated the infection of a variety of human cells that bore those surface antigens with an ecotropic virus in vitro (Roux et al., 1989).

[0310] 3. Adeno-Associated Viruses

[0311] AAV (Ridgeway, 1988; Hermonat and Muzycska, 1984) is a parovirus, discovered as a contamination of adenoviral stocks. It is a ubiquitous virus (antibodies are present in 85% of the US human population) that has not been linked to any disease. It is also classified as a dependovirus, because its replications is dependent on the presence of a helper virus, such as adenovirus. Five serotypes have been isolated, of which MV-2 is the best characterized. MV has a single-stranded linear DNA that is encapsidated into capsid proteins VP1, VP2 and VP3 to form an icosahedral virion of 20 to 24 nm in diameter (Muzyczka and McLaughlin, 1988).

[0312] The MV DNA is approximately 4.7 kilobases long. It contains two open reading frames and is flanked by two ITRs (FIG. 2). There are two major genes in the MV genome: rep and cap. The rep gene codes for proteins responsible for viral replications, whereas cap codes for capsid protein VP1-3. Each ITR forms a T-shaped hairpin structure. These terminal repeats are the only essential cis components of the AAV for chromosomal integration. Therefore, the AAV can be used as a vector with all viral coding sequences removed and replaced by the cassette of genes for delivery. Three viral promoters have been identified and named p5, p19, and p40, according to their map position. Transcription from p5 and p19 results in production of rep proteins, and transcription from p40 produces the capsid proteins (Hermonat and Muzyczka, 1984).

[0313] There are several factors that prompted researchers to study the possibility of using rAAV as an expression vector. One is that the requirements for delivering a gene to integrate into the host chromosome are surprisingly few. It is necessary to have the 145-bp ITRs, which are only 6% of the AAV genome. This leaves room in the vector to assemble a 4.5-kb DNA insertion. While this carrying capacity may prevent the AAV from delivering large genes, it is amply suited for delivering the antisense constructs of the present invention.

[0314] AAV is also a good choice of delivery vehicles due to its safety. There is a relatively complicated rescue mechanism: not only wild type adenovirus but also AAV genes are required to mobilize rAAV. Likewise, AAV is not pathogenic and not associated with any disease. The removal of viral coding sequences minimizes immune reactions to viral gene expression, and therefore, rAAV does not evoke an inflammatory response.

[0315] 4. Other Viral Vectors as Expression Constructs

[0316] Other viral vectors may be employed as expression constructs in the present invention for the delivery of oligonucleotide or polynucleotide sequences to a host cell. Vectors derived from viruses such as vaccinia virus (Ridgeway, 1988; Coupar et al., 1988), lentiviruses, polio viruses and herpes viruses may be employed. They offer several attractive features for various mammalian cells (Friedmann, 1989; Ridgeway, 1988; Coupar et al., 1988; Horwich et al., 1990).

[0317] With the recent recognition of defective hepatitis B viruses, new insight was gained into the structure-function relationship of different viral sequences. In vitro studies showed that the virus could retain the ability for helper-dependent packaging and reverse transcription despite the deletion of up to 80% of its genome (Horwich et al., 1990). This suggested that large portions of the genome could be replaced with foreign genetic material. The hepatotropism and persistence (integration) were particularly attractive properties for liver-directed gene transfer. Chang et al. (1991) introduced the chloramphenicol acetyltransferase (CAT) gene into duck hepatitis B virus genome in the place of the polymerase, surface, and pre-surface coding sequences. It was co-transfected with wild-type virus into an avian hepatoma cell line. Culture media containing high titers of the recombinant virus were used to infect primary duckling hepatocytes. Stable CAT gene expression was detected for at least 24 days after transfection (Chang et al., 1991).

[0318] 5. Non-Viral Vectors

[0319] In order to effect expression of the oligonucleotide or polynucleotide sequences of the present invention, the expression construct must be delivered into a cell. This delivery may be accomplished in vitro, as in laboratory procedures for transforming cells lines, or in vivo or ex vivo, as in the treatment of certain disease states. As described above, one preferred mechanism for delivery is via viral infection where the expression construct is encapsulated in an infectious viral particle.

[0320] Once the expression construct has been delivered into the cell the nucleic acid encoding the desired oligonucleotide or polynucleotide sequences may be positioned and expressed at different sites. In certain embodiments, the nucleic acid encoding the construct may be stably integrated into the genome of the cell. This integration may be in the specific location and orientation via homologous recombination (gene replacement) or it may be integrated in a random, non-specific location (gene augmentation). In yet further embodiments, the nucleic acid may be stably maintained in the cell as a separate, episomal segment of DNA. Such nucleic acid segments or “episomes” encode sequences sufficient to permit maintenance and replication independent of or in synchronization with the host cell cycle. How the expression construct is delivered to a cell and where in the cell the nucleic acid remains is dependent on the type of expression construct employed.

[0321] In certain embodiments of the invention, the expression construct comprising one or more oligonucleotide or polynucleotide sequences may simply consist of naked recombinant DNA or plasmids. Transfer of the construct may be performed by any of the methods mentioned above which physically or chemically permeabilize the cell membrane. This is particularly applicable for transfer in vitro but it may be applied to in vivo use as well. Dubensky et al. (1984) successfully injected polyomavirus DNA in the form of calcium phosphate precipitates into liver and spleen of adult and newborn mice demonstrating active viral replication and acute infection. Benvenisty and Reshef (1986) also demonstrated that direct intraperitoneal injection of calcium phosphate-precipitated plasmids results in expression of the transfected genes. It is envisioned that DNA encoding a gene of interest may also be transferred in a similar manner in vivo and express the gene product.

[0322] Another embodiment of the invention for transferring a naked DNA expression construct into cells may involve particle bombardment. This method depends on the ability to accelerate DNA-coated microprojectiles to a high velocity allowing them to pierce cell membranes and enter cells without killing them (Klein et al., 1987). Several devices for accelerating small particles have been developed. One such device relies on a high voltage discharge to generate an electrical current, which in turn provides the motive force (Yang et al., 1990). The microprojectiles used have consisted of biologically inert substances such as tungsten or gold beads.

[0323] Selected organs including the liver, skin, and muscle tissue of rats and mice have been bombarded in vivo (Yang et al., 1990; Zelenin et al., 1991). This may require surgical exposure of the tissue or cells, to eliminate any intervening tissue between the gun and the target organ, i.e., ex vivo treatment. Again, DNA encoding a particular gene may be delivered via this method and still be incorporated by the present invention.

[0324] Antisense Oligonucleotides

[0325] The end result of the flow of genetic information is the synthesis of protein. DNA is transcribed by polymerases into messenger RNA and translated on the ribosome to yield a folded, functional protein. Thus there are several steps along the route where protein synthesis can be inhibited. The native DNA segment coding for a polypeptide described herein, as all such mammalian DNA strands, has two strands: a sense strand and an antisense strand held together by hydrogen bonding. The messenger RNA coding for polypeptide has the same nucleotide sequence as the sense DNA strand except that the DNA thymidine is replaced by uridine. Thus, synthetic antisense nucleotide sequences will bind to a mRNA and inhibit expression of the protein encoded by that mRNA.

[0326] The targeting of antisense oligonucleotides to mRNA is thus one mechanism to shut down protein synthesis, and, consequently, represents a powerful and targeted therapeutic approach. For example, the synthesis of polygalactauronase and the muscarine type 2 acetylcholine receptor are inhibited by antisense oligonucleotides directed to their respective mRNA sequences (U.S. Pat. No. 5,739,119 and U.S. Pat. No. 5,759,829, each specifically incorporated herein by reference in its entirety). Further, examples of antisense inhibition have been demonstrated with the nuclear protein cyclin, the multiple drug resistance gene (MDG1), ICAM-1, E-selectin, STK-1, striatal GABAA receptor and human EGF (Jaskulski et al., 1988; Vasanthakumar and Ahmed, 1989; Peris et al., 1998; U.S. Pat. No. 5,801,154; U.S. Pat. No. 5,789,573; U.S. Pat. No. 5,718,709 and U.S. Pat. No. 5,610,288, each specifically incorporated herein by reference in its entirety). Antisense constructs have also been described that inhibit and can be used to treat a variety of abnormal cellular proliferations, e.g. cancer (U.S. Pat. No. 5,747,470; U.S. Pat. No. 5,591,317 and U.S. Pat. No. 5,783,683, each specifically incorporated herein by reference in its entirety).

[0327] Therefore, in exemplary embodiments, the invention provides oligonucleotide sequences that comprise all, or a portion of, any sequence that is capable of specifically binding to polynucleotide sequence described herein, or a complement thereof. In one embodiment, the antisense oligonucleotides comprise DNA or derivatives thereof. In another embodiment, the oligonucleotides comprise RNA or derivatives thereof. In a third embodiment, the oligonucleotides are modified DNAs comprising a phosphorothioated modified backbone. In a fourth embodiment, the oligonucleotide sequences comprise peptide nucleic acids or derivatives thereof. In each case, preferred compositions comprise a sequence region that is complementary, and more preferably substantially-complementary, and even more preferably, completely complementary to one or more portions of polynucleotides disclosed herein.

[0328] Selection of antisense compositions specific for a given gene sequence is based upon analysis of the chosen target sequence (i.e., in these illustrative examples the rat and human sequences) and determination of secondary structure, T_(m), binding energy, relative stability, and antisense compositions were selected based upon their relative inability to form dimers, hairpins, or other secondary structures that would reduce or prohibit specific binding to the target mRNA in a host cell.

[0329] Highly preferred target regions of the mRNA, are those which are at or near the AUG translation initiation codon, and those sequences which were substantially complementary to 5′ regions of the mRNA. These secondary structure analyses and target site selection considerations were performed using v.4 of the OLIGO primer analysis software (Rychlik, 1997) and the BLASTN 2.0.5 algorithm software (Altschul et al., 1997).

[0330] The use of an antisense delivery method employing a short peptide vector, termed MPG (27 residues), is also contemplated. The MPG peptide contains a hydrophobic domain derived from the fusion sequence of HIV gp41 and a hydrophilic domain from the nuclear localization sequence of SV40 T-antigen (Morris et al., 1997). It has been demonstrated that several molecules of the MPG peptide coat the antisense oligonucleotides and can be delivered into cultured mammalian cells in less than 1 hour with relatively high efficiency (90%). Further, the interaction with MPG strongly increases both the stability of the oligonucleotide to nuclease and the ability to cross the plasma membrane (Morris et al., 1997).

[0331] Ribozymes

[0332] Although proteins traditionally have been used for catalysis of nucleic acids, another class of macromolecules has emerged as useful in this endeavor. Ribozymes are RNA-protein complexes that cleave nucleic acids in a site-specific fashion. Ribozymes have specific catalytic domains that possess endonuclease activity (Kim and Cech, 1987; Gerlach et al., 1987; Forster and Symons, 1987). For example, a large number of ribozymes accelerate phosphoester transfer reactions with a high degree of specificity, often cleaving only one of several phosphoesters in an oligonucleotide substrate (Cech et al., 1981; Michel and Westhof, 1990; Reinhold-Hurek and Shub, 1992). This specificity has been attributed to the requirement that the substrate bind via specific base-pairing interactions to the internal guide sequence (“IGS”) of the ribozyme prior to chemical reaction.

[0333] Ribozyme catalysis has primarily been observed as part of sequence-specific cleavage/ligation reactions involving nucleic acids (Joyce, 1989; Cech et al., 1981). For example, U.S. Pat. No. 5,354,855 (specifically incorporated herein by reference) reports that certain ribozymes can act as endonucleases with a sequence specificity greater than that of known ribonucleases and approaching that of the DNA restriction enzymes. Thus, sequence-specific ribozyme-mediated inhibition of gene expression may be particularly suited to therapeutic applications (Scanlon et al., 1991; Sarver et al., 1990). Recently, it was reported that ribozymes elicited genetic changes in some cells lines to which they were applied; the altered genes included the oncogenes H-ras, c-fos and genes of HIV. Most of this work involved the modification of a target mRNA, based on a specific mutant codon that is cleaved by a specific ribozyme.

[0334] Six basic varieties of naturally-occurring enzymatic RNAs are known presently. Each can catalyze the hydrolysis of RNA phosphodiester bonds in trans (and thus can cleave other RNA molecules) under physiological conditions. In general, enzymatic nucleic acids act by first binding to a target RNA. Such binding occurs through the target binding portion of a enzymatic nucleic acid which is held in close proximity to an enzymatic portion of the molecule that acts to cleave the target RNA. Thus, the enzymatic nucleic acid first recognizes and then binds a target RNA through complementary base-pairing, and once bound to the correct site, acts enzymatically to cut the target RNA. Strategic cleavage of such a target RNA will destroy its ability to direct synthesis of an encoded protein. After an enzymatic nucleic acid has bound and cleaved its RNA target, it is released from that RNA to search for another target and can repeatedly bind and cleave new targets.

[0335] The enzymatic nature of a ribozyme is advantageous over many technologies, such as antisense technology (where a nucleic acid molecule simply binds to a nucleic acid target to block its translation) since the concentration of ribozyme necessary to affect a therapeutic treatment is lower than that of an antisense oligonucleotide. This advantage reflects the ability of the ribozyme to act enzymatically. Thus, a single ribozyme molecule is able to cleave many molecules of target RNA. In addition, the ribozyme is a highly specific inhibitor, with the specificity of inhibition depending not only on the base pairing mechanism of binding to the target RNA, but also on the mechanism of target RNA cleavage. Single mismatches, or base-substitutions, near the site of cleavage can completely eliminate catalytic activity of a ribozyme. Similar mismatches in antisense molecules do not prevent their action (Woolf et al., 1992). Thus, the specificity of action of a ribozyme is greater than that of an antisense oligonucleotide binding the same RNA site.

[0336] The enzymatic nucleic acid molecule may be formed in a hammerhead, hairpin, a hepatitis δ virus, group I intron or RNaseP RNA (in association with an RNA guide sequence) or Neurospora VS RNA motif. Examples of hammerhead motifs are described by Rossi et al. (1992). Examples of hairpin motifs are described by Hampel et al. (Eur. Pat. Appl. Publ. No. EP 0360257), Hampel and Tritz (1989), Hampel et al. (1990) and U.S. Pat. No. 5,631,359 (specifically incorporated herein by reference). An example of the hepatitis δ virus motif is described by Perrotta and Been (1992); an example of the RNaseP motif is described by Guerrier-Takada et al. (1983); Neurospora VS RNA ribozyme motif is described by Collins (Saville and Collins, 1990; Saville and Collins, 1991; Collins and Olive, 1993); and an example of the Group I intron is described in (U.S. Pat. No. 4,987,071, specifically incorporated herein by reference). All that is important in an enzymatic nucleic acid molecule of this invention is that it has a specific substrate binding site which is complementary to one or more of the target gene RNA regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart an RNA cleaving activity to the molecule. Thus the ribozyme constructs need not be limited to specific motifs mentioned herein.

[0337] In certain embodiments, it may be important to produce enzymatic cleaving agents which exhibit a high degree of specificity for the RNA of a desired target, such as one of the sequences disclosed herein. The enzymatic nucleic acid molecule is preferably targeted to a highly conserved sequence region of a target mRNA. Such enzymatic nucleic acid molecules can be delivered exogenously to specific cells as required. Alternatively, the ribozymes can be expressed from DNA or RNA vectors that are delivered to specific cells.

[0338] Small enzymatic nucleic acid motifs (e.g., of the hammerhead or the hairpin structure) may also be used for exogenous delivery. The simple structure of these molecules increases the ability of the enzymatic nucleic acid to invade targeted regions of the mRNA structure. Alternatively, catalytic RNA molecules can be expressed within cells from eukaryotic promoters (e.g., Scanlon et al., 1991; Kashani-Sabet et al., 1992; Dropulic et al., 1992; Weerasinghe et al., 1991; Ojwang et al., 1992; Chen et al., 1992; Sarver et al., 1990). Those skilled in the art realize that any ribozyme can be expressed in eukaryotic cells from the appropriate DNA vector. The activity of such ribozymes can be augmented by their release from the primary transcript by a second ribozyme (Int. Pat. Appl. Publ. No. WO 93/23569, and Int. Pat. Appl. Publ. No. WO 94/02595, both hereby incorporated by reference; Ohkawa et al., 1992; Taira et al., 1991; and Ventura et al., 1993).

[0339] Ribozymes may be added directly, or can be complexed with cationic lipids, lipid complexes, packaged within liposomes, or otherwise delivered to target cells. The RNA or RNA complexes can be locally administered to relevant tissues ex vivo, or in vivo through injection, aerosol inhalation, infusion pump or stent, with or without their incorporation in biopolymers.

[0340] Ribozymes may be designed as described in Int. Pat. Appl. Publ. No. WO 93/23569 and Int. Pat. Appl. Publ. No. WO 94/02595, each specifically incorporated herein by reference) and synthesized to be tested in vitro and in vivo, as described. Such ribozymes can also be optimized for delivery. While specific examples are provided, those in the art will recognize that equivalent RNA targets in other species can be utilized when necessary.

[0341] Hammerhead or hairpin ribozymes may be individually analyzed by computer folding (Jaeger et al., 1989) to assess whether the ribozyme sequences fold into the appropriate secondary structure. Those ribozymes with unfavorable intramolecular interactions between the binding arms and the catalytic core are eliminated from consideration. Varying binding arm lengths can be chosen to optimize activity. Generally, at least 5 or so bases on each arm are able to bind to, or otherwise interact with, the target RNA.

[0342] Ribozymes of the hammerhead or hairpin motif may be designed to anneal to various sites in the mRNA message, and can be chemically synthesized. The method of synthesis used follows the procedure for normal RNA synthesis as described in Usman et al. (1987) and in Scaringe et al. (1990) and makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. Average stepwise coupling yields are typically >98%. Hairpin ribozymes may be synthesized in two parts and annealed to reconstruct an active ribozyme (Chowrira and Burke, 1992). Ribozymes may be modified extensively to enhance stability by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-flouro, 2′-o-methyl, 2′-H (for a review see, e.g., Usman and Cedergren, 1992). Ribozymes may be purified by gel electrophoresis using general methods or by high pressure liquid chromatography and resuspended in water.

[0343] Ribozyme activity can be optimized by altering the length of the ribozyme binding arms, or chemically synthesizing ribozymes with modifications that prevent their degradation by serum ribonucleases (see, e.g., Int. Pat. Appl. Publ. No. WO 92/07065; Perrault et al, 1990; Pieken et al., 1991; Usman and Cedergren, 1992; Int. Pat. Appl. Publ. No. WO 93/15187; Int. Pat. Appl. Publ. No. WO 91/03162; Eur. Pat. Appl. Publ. No. 92110298.4; U.S. Pat. No. 5,334,711; and Int. Pat. Appl. Publ. No. WO 94/13688, which describe various chemical modifications that can be made to the sugar moieties of enzymatic RNA molecules), modifications which enhance their efficacy in cells, and removal of stem II bases to shorten RNA synthesis times and reduce chemical requirements.

[0344] Sullivan et al. (Int. Pat. Appl. Publ. No. WO 94/02595) describes the general methods for delivery of enzymatic RNA molecules. Ribozymes may be administered to cells by a variety of methods known to those familiar to the art, including, but not restricted to, encapsulation in liposomes, by iontophoresis, or by incorporation into other vehicles, such as hydrogels, cyclodextrins, biodegradable nanocapsules, and bioadhesive microspheres. For some indications, ribozymes may be directly delivered ex vivo to cells or tissues with or without the aforementioned vehicles. Alternatively, the RNA/vehicle combination may be locally delivered by direct inhalation, by direct injection or by use of a catheter, infusion pump or stent. Other routes of delivery include, but are not limited to, intravascular, intramuscular, subcutaneous or joint injection, aerosol inhalation, oral (tablet or pill form), topical, systemic, ocular, intraperitoneal and/or intrathecal delivery. More detailed descriptions of ribozyme delivery and administration are provided in Int. Pat. Appl. Publ. No. WO 94/02595 and Int. Pat. Appl. Publ. No. WO 93/23569, each specifically incorporated herein by reference.

[0345] Another means of accumulating high concentrations of a ribozyme(s) within cells is to incorporate the ribozyme-encoding sequences into a DNA expression vector. Transcription of the ribozyme sequences are driven from a promoter for eukaryotic RNA polymerase I (pol I), RNA polymerase II (pol II), or RNA polymerase III (pol III). Transcripts from pol II or pol III promoters will be expressed at high levels in all cells; the levels of a given pol II promoter in a given cell type will depend on the nature of the gene regulatory sequences (enhancers, silencers, etc.) present nearby. Prokaryotic RNA polymerase promoters may also be used, providing that the prokaryotic RNA polymerase enzyme is expressed in the appropriate cells (Elroy-Stein and Moss, 1990; Gao and Huang, 1993; Lieber et al., 1993; Zhou et al., 1990). Ribozymes expressed from such promoters can function in mammalian cells (e.g., Kashani-Saber et al., 1992; Ojwang et al., 1992; Chen et al., 1992; Yu et al., 1993; L′Huillier et al., 1992; Lisziewicz et al., 1993). Such transcription units can be incorporated into a variety of vectors for introduction into mammalian cells, including but not restricted to, plasmid DNA vectors, viral DNA vectors (such as adenovirus or adeno-associated vectors), or viral RNA vectors (such as retroviral, semliki forest virus, sindbis virus vectors).

[0346] Ribozymes may be used as diagnostic tools to examine genetic drift and mutations within diseased cells. They can also be used to assess levels of the target RNA molecule. The close relationship between ribozyme activity and the structure of the target RNA allows the detection of mutations in any region of the molecule which alters the base-pairing and three-dimensional structure of the target RNA. By using multiple ribozymes, one may map nucleotide changes which are important to RNA structure and function in vitro, as well as in cells and tissues. Cleavage of target RNAs with ribozymes may be used to inhibit gene expression and define the role (essentially) of specified gene products in the progression of disease. In this manner, other genetic targets may be defined as important mediators of the disease. These studies will lead to better treatment of the disease progression by affording the possibility of combinational therapies (e.g., multiple ribozymes targeted to different genes, ribozymes coupled with known small molecule inhibitors, or intermittent treatment with combinations of ribozymes and/or other chemical or biological molecules). Other in vitro uses of ribozymes are well known in the art, and include detection of the presence of mRNA associated with an IL-5 related condition. Such RNA is detected by determining the presence of a cleavage product after treatment with a ribozyme using standard methodology.

[0347] Peptide Nucleic Acids

[0348] In certain embodiments, the inventors contemplate the use of peptide nucleic acids (PNAs) in the practice of the methods of the invention. PNA is a DNA mimic in which the nucleobases are attached to a pseudopeptide backbone (Good and Nielsen, 1997). PNA is able to be utilized in a number methods that traditionally have used RNA or DNA. Often PNA sequences perform better in techniques than the corresponding RNA or DNA sequences and have utilities that are not inherent to RNA or DNA. A review of PNA including methods of making, characteristics of, and methods of using, is provided by Corey (1997) and is incorporated herein by reference. As such, in certain embodiments, one may prepare PNA sequences that are complementary to one or more portions of the ACE mRNA sequence, and such PNA compositions may be used to regulate, alter, decrease, or reduce the translation of ACE-specific mRNA, and thereby alter the level of ACE activity in a host cell to which such PNA compositions have been administered.

[0349] PNAs have 2-aminoethyl-glycine linkages replacing the normal phosphodiester backbone of DNA (Nielsen et al., 1991; Hanvey et al., 1992; Hyrup and Nielsen, 1996; Neilsen, 1996). This chemistry has three important consequences: firstly, in contrast to DNA or phosphorothioate oligonucleotides, PNAs are neutral molecules; secondly, PNAs are achiral, which avoids the need to develop a stereoselective synthesis; and thirdly, PNA synthesis uses standard Boc (Dueholm et al., 1994) or Fmoc (Thomson et al., 1995) protocols for solid-phase peptide synthesis, although other methods, including a modified Merrifield method, have been used (Christensen et al., 1995).

[0350] PNA monomers or ready-made oligomers are commercially available from PerSeptive Biosystems (Framingham, MA). PNA syntheses by either Boc or Fmoc protocols are straightforward using manual or automated protocols (Norton et al., 1995). The manual protocol lends itself to the production of chemically modified PNAs or the simultaneous synthesis of families of closely related PNAs.

[0351] As with peptide synthesis, the success of a particular PNA synthesis will depend on the properties of the chosen sequence. For example, while in theory PNAs can incorporate any combination of nucleotide bases, the presence of adjacent purines can lead to deletions of one or more residues in the product. In expectation of this difficulty, it is suggested that, in producing PNAs with adjacent purines, one should repeat the coupling of residues likely to be added inefficiently. This should be followed by the purification of PNAs by reverse-phase high-pressure liquid chromatography (Norton et al., 1995) providing yields and purity of product similar to those observed during the synthesis of peptides.

[0352] Modifications of PNAs for a given application may be accomplished by coupling amino acids during solid-phase synthesis or by attaching compounds that contain a carboxylic acid group to the exposed N-terminal amine. Alternatively, PNAs can be modified after synthesis by coupling to an introduced lysine or cysteine. The ease with which PNAs can be modified facilitates optimization for better solubility or for specific functional requirements. Once synthesized, the identity of PNAs and their derivatives can be confirmed by mass spectrometry. Several studies have made and utilized modifications of PNAs (Norton et al., 1995; Haaima et al., 1996; Stetsenko et al., 1996; Petersen et al., 1995; Ulmann et al., 1996; Koch et al., 1995; Orum et al., 1995; Footer et al., 1996; Griffith et al., 1995; Kremsky et al., 1996; Pardridge et al., 1995; Boffa et al., 1995; Landsdorp et al., 1996; Gambacorti-Passerini et al., 1996; Armitage et al., 1997; Seeger et al., 1997; Ruskowski et al., 1997). U.S. Pat. No. 5,700,922 discusses PNA-DNA-PNA chimeric molecules and their uses in diagnostics, modulating protein in organisms, and treatment of conditions susceptible to therapeutics.

[0353] In contrast to DNA and RNA, which contain negatively charged linkages, the PNA backbone is neutral. In spite of this dramatic alteration, PNAs recognize complementary DNA and RNA by Watson-Crick pairing (Egholm et al., 1993), validating the initial modeling by Nielsen et al. (1991). PNAs lack 3′ to 5′ polarity and can bind in either parallel or antiparallel fashion, with the antiparallel mode being preferred (Egholm et al., 1993).

[0354] Hybridization of DNA oligonucleotides to DNA and RNA is destabilized by electrostatic repulsion between the negatively charged phosphate backbones of the complementary strands. By contrast, the absence of charge repulsion in PNA-DNA or PNA-RNA duplexes increases the melting temperature (T_(m)) and reduces the dependence of Tm on the concentration of mono- or divalent cations (Nielsen et al., 1991). The enhanced rate and affinity of hybridization are significant because they are responsible for the surprising ability of PNAs to perform strand invasion of complementary sequences within relaxed double-stranded DNA. In addition, the efficient hybridization at inverted repeats suggests that PNAs can recognize secondary structure effectively within double-stranded DNA. Enhanced recognition also occurs with PNAs immobilized on surfaces, and Wang et al. have shown that support-bound PNAs can be used to detect hybridization events (Wang et al., 1996).

[0355] One might expect that tight binding of PNAs to complementary sequences would also increase binding to similar (but not identical) sequences, reducing the sequence specificity of PNA recognition. As with DNA hybridization, however, selective recognition can be achieved by balancing oligomer length and incubation temperature. Moreover, selective hybridization of PNAs is encouraged by PNA-DNA hybridization being less tolerant of base mismatches than DNA-DNA hybridization. For example, a single mismatch within a 16 bp PNA-DNA duplex can reduce the Tm by up to 150C (Egholm et al., 1993). This high level of discrimination has allowed the development of several PNA-based strategies for the analysis of point mutations (Wang et al., 1996; Carlsson et al., 1996; Thiede et al., 1996; Webb and Hurskainen, 1996; Perry-O'Keefe et al., 1996).

[0356] High-affinity binding provides clear advantages for molecular recognition and the development of new applications for PNAs. For example, 11-13 nucleotide PNAs inhibit the activity of telomerase, a ribonucleo-protein that extends telomere ends using an essential RNA template, while the analogous DNA oligomers do not (Norton et al., 1996).

[0357] Neutral PNAs are more hydrophobic than analogous DNA oligomers, and this can lead to difficulty solubilizing them at neutral pH, especially if the PNAs have a high purine content or if they have the potential to form secondary structures. Their solubility can be enhanced by attaching one or more positive charges to the PNA termini (Nielsen et al., 1991).

[0358] Findings by Allfrey and colleagues suggest that strand invasion will occur spontaneously at sequences within chromosomal DNA (Boffa et al., 1995; Boffa et al., 1996). These studies targeted PNAs to triplet repeats of the nucleotides CAG and used this recognition to purify transcriptionally active DNA (Boffa et al., 1995) and to inhibit transcription (Boffa et al., 1996). This result suggests that if PNAs can be delivered within cells then they will have the potential to be general sequence-specific regulators of gene expression. Studies and reviews concerning the use of PNAs as antisense and anti-gene agents include Nielsen et al. (1993b), Hanvey et al. (1992), and Good and Nielsen (1997). Koppelhus et al. (1997) have used PNAs to inhibit HIV-1 inverse transcription, showing that PNAs may be used for antiviral therapies.

[0359] Methods of characterizing the antisense binding properties of PNAs are discussed in Rose (1993) and Jensen et al. (1997). Rose uses capillary gel electrophoresis to determine binding of PNAs to their complementary oligonucleotide, measuring the relative binding kinetics and stoichiometry. Similar types of measurements were made by Jensen et al. using BIAcore™ technology.

[0360] Other applications of PNAs include use in DNA strand invasion (Nielsen et al., 1991), antisense inhibition (Hanvey et al., 1992), mutational analysis (Orum et al., 1993), enhancers of transcription (Mollegaard et al., 1994), nucleic acid purification (Orum et al., 1995), isolation of transcriptionally active genes (Boffa et al., 1995), blocking of transcription factor binding (Vickers et al., 1995), genome cleavage (Veselkov et al., 1996), biosensors (Wang et al., 1996), in situ hybridization (Thisted et al., 1996), and in a alternative to Southern blotting (Perry-O'Keefe, 1996).

[0361] Polypeptide Compositions

[0362] The present invention, in other aspects, provides polypeptide compositions. Generally, a polypeptide of the invention will be an isolated polypeptide (or an epitope, variant, or active fragment thereof) derived from HSV. Preferably, the polypeptide is encoded by a polynucleotide sequence disclosed herein or a sequence which hybridizes under moderate or highly stringent conditions to a polynucleotide sequence disclosed herein. Alternatively, the polypeptide may be defined as a polypeptide which comprises a contiguous amino acid sequence from an amino acid sequence disclosed herein, or which polypeptide comprises an entire amino acid sequence disclosed herein.

[0363] In the present invention, a polypeptide composition is also understood to comprise one or more polypeptides that are immunologically reactive with antibodies and/or T cells generated against a polypeptide of the invention, particularly a polypeptide having amino acid sequences disclosed herein, or to active fragments, or to variants or biological functional equivalents thereof.

[0364] Likewise, a polypeptide composition of the present invention is understood to comprise one or more polypeptides that are capable of eliciting antibodies or T cells that are immunologically reactive with one or more polypeptides encoded by one or more contiguous nucleic acid sequences contained in the amino acid sequences disclosed herein, or to active fragments, or to variants thereof, or to one or more nucleic acid sequences which hybridize to one or more of these sequences under conditions of moderate to high stringency. Particularly illustrative polypeptides comprise the amino acid sequence disclosed in SEQ ID NO: 2, 3, 5, 6, 7, 10-12, 14-15, 17-18, 20-23, 25-33, 39-47, 50-51, 54-64, 74-75, 90-97, 120-121, 122-140, 142-143, 153-178, and 181.

[0365] As used herein, an active fragment of a polypeptide includes a whole or a portion of a polypeptide which is modified by conventional techniques, e.g., mutagenesis, or by addition, deletion, or substitution, but which active fragment exhibits substantially the same structure function, antigenicity, etc., as a polypeptide as described herein.

[0366] In certain illustrative embodiments, the polypeptides of the invention will comprise at least an immunogenic portion of an HSV antigen or a variant or biological functional equivalent thereof, as described herein. Polypeptides as described herein may be of any length. Additional sequences derived from the native protein and/or heterologous sequences may be present, and such sequences may (but need not) possess further immunogenic or antigenic properties.

[0367] An “immunogenic portion,” as used herein is a portion of a protein that is recognized (i.e., specifically bound) by a B-cell and/or T-cell surface antigen receptor. Such immunogenic portions generally comprise at least 5 amino acid residues, more preferably at least 10, and still more preferably at least 20 amino acid residues of an HSV protein or a variant thereof. Certain preferred immunogenic portions include peptides in which an N-terminal leader sequence and/or transmembrane domain have been deleted. Other preferred immunogenic portions may contain a small N- and/or C-terminal deletion (e.g., 1-30 amino acids, preferably 5-15 amino acids), relative to the mature protein.

[0368] Immunogenic portions may generally be identified using well known techniques, such as those summarized in Paul, Fundamental Immunology, 3rd ed., 243-247 (Raven Press, 1993) and references cited therein. Such techniques include screening polypeptides for the ability to react with antigen-specific antibodies, antisera and/or T-cell lines or clones. As used herein, antisera and antibodies are “antigen-specific” if they specifically bind to an antigen (i.e., they react with the protein in an ELISA or other immunoassay, and do not react detectably with unrelated proteins). Such antisera and antibodies may be prepared as described herein, and using well known techniques. An immunogenic portion of a native HSV protein is a portion that reacts with such antisera and/or T-cells at a level that is not substantially less than the reactivity of the full length polypeptide (e.g., in an ELISA and/or T-cell reactivity assay). Such immunogenic portions may react within such assays at a level that is similar to or greater than the reactivity of the full length polypeptide. Such screens may generally be performed using methods well known to those of ordinary skill in the art, such as those described in Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988. For example, a polypeptide may be immobilized on a solid support and contacted with patient sera to allow binding of antibodies within the sera to the immobilized polypeptide. Unbound sera may then be removed and bound antibodies detected using, for example, ¹²⁵I-labeled Protein A.

[0369] As noted above, a composition may comprise a variant of a native HSV protein. A polypeptide “variant,” as used herein, is a polypeptide that differs from a native HSV protein in one or more substitutions, deletions, additions and/or insertions, such that the immunogenicity of the polypeptide is not substantially diminished. In other words, the ability of a variant to react with antigen-specific antisera may be enhanced or unchanged, relative to the native protein, or may be diminished by less than 50%, and preferably less than 20%, relative to the native protein. Such variants may generally be identified by modifying one of the above polypeptide sequences and evaluating the reactivity of the modified polypeptide with antigen-specific antibodies or antisera as described herein. Preferred variants include those in which one or more portions, such as an N-terminal leader sequence or transmembrane domain, have been removed. Other preferred variants include variants in which a small portion (e.g., 1-30 amino acids, preferably 5-15 amino acids) has been removed from the N- and/or C-terminal of the mature protein.

[0370] Polypeptide variants encompassed by the present invention include those exhibiting at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more identity (determined as described above) to the polypeptides disclosed herein.

[0371] Preferably, a variant contains conservative substitutions. A “conservative substitution” is one in which an amino acid is substituted for another amino acid that has similar properties, such that one skilled in the art of peptide chemistry would expect the secondary structure and hydropathic nature of the polypeptide to be substantially unchanged. Amino acid substitutions may generally be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity and/or the amphipathic nature of the residues. For example, negatively charged amino acids include aspartic acid and glutamic acid; positively charged amino acids include lysine and arginine; and amino acids with uncharged polar head groups having similar hydrophilicity values include leucine, isoleucine and valine; glycine and alanine; asparagine and glutamine; and serine, threonine, phenylalanine and tyrosine. Other groups of amino acids that may represent conservative changes include: (1) ala, pro, gly, glu, asp, gin, asn, ser, thr; (2) cys, ser, tyr, thr; (3) val, ile, leu, met, ala, phe; (4) lys, arg, his; and (5) phe, tyr, trp, his. A variant may also, or alternatively, contain nonconservative changes. In a preferred embodiment, variant polypeptides differ from a native sequence by substitution, deletion or addition of five amino acids or fewer. Variants may also (or alternatively) be modified by, for example, the deletion or addition of amino acids that have minimal influence on the immunogenicity, secondary structure and hydropathic nature of the polypeptide.

[0372] As noted above, polypeptides may comprise a signal (or leader) sequence at the N-terminal end of the protein, which co-translationally or post-translationally directs transfer of the protein. The polypeptide may also be conjugated to a linker or other sequence for ease of synthesis, purification or identification of the polypeptide (e.g., poly-His), or to enhance binding of the polypeptide to a solid support. For example, a polypeptide may be conjugated to an immunoglobulin Fc region.

[0373] Polypeptides may be prepared using any of a variety of well known techniques. Recombinant polypeptides encoded by DNA sequences as described above may be readily prepared from the DNA sequences using any of a variety of expression vectors known to those of ordinary skill in the art. Expression may be achieved in any appropriate host cell that has been transformed or transfected with an expression vector containing a DNA molecule that encodes a recombinant polypeptide. Suitable host cells include prokaryotes, yeast, and higher eukaryotic cells, such as mammalian cells and plant cells. Preferably, the host cells employed are E. coli, yeast or a mammalian cell line such as COS or CHO. Supernatants from suitable host/vector systems which secrete recombinant protein or polypeptide into culture media may be first concentrated using a commercially available filter. Following concentration, the concentrate may be applied to a suitable purification matrix such as an affinity matrix or an ion exchange resin. Finally, one or more reverse phase HPLC steps can be employed to further purify a recombinant polypeptide.

[0374] Portions and other variants having less than about 100 amino acids, and generally less than about 50 amino acids, may also be generated by synthetic means, using techniques well known to those of ordinary skill in the art. For example, such polypeptides may be synthesized using any of the commercially available solid-phase techniques, such as the Merrifield solid-phase synthesis method, where amino acids are sequentially added to a growing amino acid chain. See Merrifield, J. Am. Chem. Soc. 85:2149-2146, 1963. Equipment for automated synthesis of polypeptides is commercially available from suppliers such as Perkin Elmer/Applied BioSystems Division (Foster City, Calif.), and may be operated according to the manufacturer's instructions.

[0375] Within certain specific embodiments, a polypeptide may be a fusion protein that comprises multiple polypeptides as described herein, or that comprises at least one polypeptide as described herein and an unrelated sequence, such as a known protein. A fusion partner may, for example, assist in providing T helper epitopes (an immunological fusion partner), preferably T helper epitopes recognized by humans, or may assist in expressing the protein (an expression enhancer) at higher yields than the native recombinant protein. Certain preferred fusion partners are both immunological and expression enhancing fusion partners. Other fusion partners may be selected so as to increase the solubility of the protein or to enable the protein to be targeted to desired intracellular compartments. Still further fusion partners include affinity tags, which facilitate purification of the protein.

[0376] Fusion proteins may generally be prepared using standard techniques, including chemical conjugation. Preferably, a fusion protein is expressed as a recombinant protein, allowing the production of increased levels, relative to a non-fused protein, in an expression system. Briefly, DNA sequences encoding the polypeptide components may be assembled separately, and ligated into an appropriate expression vector. The 3′ end of the DNA sequence encoding one polypeptide component is ligated, with or without a peptide linker, to the 5′ end of a DNA sequence encoding the second polypeptide component so that the reading frames of the sequences are in phase. This permits translation into a single fusion protein that retains the biological activity of both component polypeptides.

[0377] A peptide linker sequence may be employed to separate the first and second polypeptide components by a distance sufficient to ensure that each polypeptide folds into its secondary and tertiary structures. Such a peptide linker sequence is incorporated into the fusion protein using standard techniques well known in the art. Suitable peptide linker sequences may be chosen based on the following factors: (1) their ability to adopt a flexible extended conformation; (2) their inability to adopt a secondary structure that could interact with functional epitopes on the first and second polypeptides; and (3) the lack of hydrophobic or charged residues that might react with the polypeptide functional epitopes. Preferred peptide linker sequences contain Gly, Asn and Ser residues. Other near neutral amino acids, such as Thr and Ala may also be used in the linker sequence. Amino acid sequences which may be usefully employed as linkers include those disclosed in Maratea et al., Gene 40:39-46,1985; Murphy et al., Proc. Natl. Acad. Sci. USA 83:8258-8262,1986; U.S. Pat. No. 4,935,233 and U.S. Pat. No. 4,751,180. The linker sequence may generally be from 1 to about 50 amino acids in length. Linker sequences are not required when the first and second polypeptides have non-essential N-terminal amino acid regions that can be used to separate the functional domains and prevent steric interference.

[0378] The ligated DNA sequences are operably linked to suitable transcriptional or translational regulatory elements. The regulatory elements responsible for expression of DNA are located only 5′ to the DNA sequence encoding the first polypeptides. Similarly, stop codons required to end translation and transcription termination signals are only present 3′ to the DNA sequence encoding the second polypeptide.

[0379] Fusion proteins are also provided. Such proteins comprise a polypeptide as described herein together with an unrelated immunogenic protein. Preferably the immunogenic protein is capable of eliciting a recall response. Examples of such proteins include tetanus, tuberculosis and hepatitis proteins (see, for example, Stoute et al. New Engl. J. Med., 336:86-91, 1997).

[0380] Within preferred embodiments, an immunological fusion partner is derived from protein D, a surface protein of the gram-negative bacterium Haemophilus influenza B (WO 91/18926). Preferably, a protein D derivative comprises approximately the first third of the protein (e.g., the first N-terminal 100-110 amino acids), and a protein D derivative may be lipidated. Within certain preferred embodiments, the first 109 residues of a Lipoprotein D fusion partner is included on the N-terminus to provide the polypeptide with additional exogenous T-cell epitopes and to increase the expression level in E. coli (thus functioning as an expression enhancer). The lipid tail ensures optimal presentation of the antigen to antigen presenting cells. Other fusion partners include the non-structural protein from influenzae virus, NS1 (hemaglutinin). Typically, the N-terminal 81 amino acids are used, although different fragments that include T-helper epitopes may be used.

[0381] In another embodiment, a Mycobacterium tuberculosis-derived Ra12 polynucleotide is linked to at least an immunogenic portion of an HSV polynucleotide of this invention. Ra12 compositions and methods for their use in enhancing expression of heterologous polynucleotide sequences is described in U.S. Patent Application No. 60/158,585, the disclosure of which is incorporated herein by reference in its entirety. Briefly, Ra12 refers to a polynucleotide region that is a subsequence of a Mycobacterium tuberculosis MTB32A nucleic acid. MTB32A is a serine protease of 32 KD molecular weight encoded by a gene in virulent and avirulent strains of M. tuberculosis. The nucleotide sequence and amino acid sequence of MTB32A have been disclosed (U.S. Patent Application No. 60/158,585; see also, Skeiky et al., Infection and Immun. (1999) 67:3998-4007, incorporated herein by reference). The Ra12 C-terminal fragment of the MTB32A coding sequence expresses at high levels on its own and remains as a soluble protein throughout the purification process. Moreover, the presence of Ra12 polypeptide fragments in a fusion polypeptide may enhance the immunogenicity of the heterologous antigenic HSV polypeptides with which Ra12 is fused. In one embodiment, the Ra12 polypeptide sequence present in a fusion polypeptide with an HSV antigen comprises some or all of amino acid residues 192 to 323 of MTB32A.

[0382] In another embodiment, the immunological fusion partner is the protein known as LYTA, or a portion thereof (preferably a C-terminal portion). LYTA is derived from Streptococcus pneumoniae, which synthesizes an N-acetyl-L-alanine amidase known as amidase LYTA (encoded by the LytA gene; Gene 43:265-292, 1986). LYTA is an autolysin that specifically degrades certain bonds in the peptidoglycan backbone. The C-terminal domain of the LYTA protein is responsible for the affinity to the choline or to some choline analogues such as DEAE. This property has been exploited for the development of E. coli C-LYTA expressing plasmids useful for expression of fusion proteins. Purification of hybrid proteins containing the C-LYTA fragment at the amino terminus has been described (see Biotechnology 10:795-798, 1992). Within a preferred embodiment, a repeat portion of LYTA may be incorporated into a fusion protein. A repeat portion is found in the C-terminal region starting at residue 178. A particularly preferred repeat portion incorporates residues 188-305.

[0383] In general, polypeptides (including fusion proteins) and polynucleotides as described herein are isolated. An “isolated” polypeptide or polynucleotide is one that is removed from its original environment. For example, a naturally-occurring protein is isolated if it is separated from some or all of the coexisting materials in the natural system. Preferably, such polypeptides are at least about 90% pure, more preferably at least about 95% pure and most preferably at least about 99% pure. A polynucleotide is considered to be isolated if, for example, it is cloned into a vector that is not a part of the natural environment.

[0384] Binding Agents

[0385] The present invention further provides agents, such as antibodies and antigen-binding fragments thereof, that specifically bind to a HSV protein. As used herein, an antibody, or antigen-binding fragment thereof, is said to “specifically bind” to a HSV protein if it reacts at a detectable level (within, for example, an ELISA) with a HSV protein, and does not react detectably with unrelated proteins under similar conditions. As used herein, “binding” refers to a noncovalent association between two separate molecules such that a complex is formed. The ability to bind may be evaluated by, for example, determining a binding constant for the formation of the complex. The binding constant is the value obtained when the concentration of the complex is divided by the product of the component concentrations. In general, two compounds are said to “bind,” in the context of the present invention, when the binding constant for complex formation exceeds about 10³ L/mol. The binding constant may be determined using methods well known in the art.

[0386] Binding agents may be further capable of differentiating between patients with and without HSV infection using the representative assays provided herein. For example, preferably, antibodies or other binding agents that bind to a HSV protein will generate a signal indicating the presence of infection in at least about 20% of patients with the disease, and will generate a negative signal indicating the absence of the disease in at least about 90% of individuals without an HSV infection. To determine whether a binding agent satisfies this requirement, biological samples (e.g., blood, sera, sputum, urine and/or biopsies) from patients with and without HSV (as determined using standard clinical tests) may be assayed as described herein for the presence of polypeptides that bind to the binding agent. It will be apparent that a statistically significant number of samples with and without the disease should be assayed. Each binding agent should satisfy the above criteria; however, those of ordinary skill in the art will recognize that binding agents may be used in combination to improve sensitivity.

[0387] Any agent that satisfies the above requirements may be a binding agent. For example, a binding agent may be a ribosome, with or without a peptide component, an RNA molecule or a polypeptide. In a preferred embodiment, a binding agent is an antibody or an antigen-binding fragment thereof. Antibodies may be prepared by any of a variety of techniques known to those of ordinary skill in the art. See, e.g., Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988. In general, antibodies can be produced by cell culture techniques, including the generation of monoclonal antibodies as described herein, or via transfection of antibody genes into suitable bacterial or mammalian cell hosts, in order to allow for the production of recombinant antibodies. In one technique, an immunogen comprising the polypeptide is initially injected into any of a wide variety of mammals (e.g., mice, rats, rabbits, sheep or goats). In this step, the polypeptides of this invention may serve as the immunogen without modification. Alternatively, particularly for relatively short polypeptides, a superior immune response may be elicited if the polypeptide is joined to a carrier protein, such as bovine serum albumin or keyhole limpet hemocyanin. The immunogen is injected into the animal host, preferably according to a predetermined schedule incorporating one or more booster immunizations, and the animals are bled periodically. Polyclonal antibodies specific for the polypeptide may then be purified from such antisera by, for example, affinity chromatography using the polypeptide coupled to a suitable solid support.

[0388] Monoclonal antibodies specific for an antigenic polypeptide of interest may be prepared, for example, using the technique of Kohler and Milstein, Eur. J. Immunol. 6:511-519,1976, and improvements thereto. Briefly, these methods involve the preparation of immortal cell lines capable of producing antibodies having the desired specificity (i.e., reactivity with the polypeptide of interest). Such cell lines may be produced, for example, from spleen cells obtained from an animal immunized as described above. The spleen cells are then immortalized by, for example, fusion with a myeloma cell fusion partner, preferably one that is syngeneic with the immunized animal. A variety of fusion techniques may be employed. For example, the spleen cells and myeloma cells may be combined with a nonionic detergent for a few minutes and then plated at low density on a selective medium that supports the growth of hybrid cells, but not myeloma cells. A preferred selection technique uses HAT (hypoxanthine, aminopterin, thymidine) selection. After a sufficient time, usually about 1 to 2 weeks, colonies of hybrids are observed. Single colonies are selected and their culture supernatants tested for binding activity against the polypeptide. Hybridomas having high reactivity and specificity are preferred.

[0389] Monoclonal antibodies may be isolated from the supernatants of growing hybridoma colonies. In addition, various techniques may be employed to enhance the yield, such as injection of the hybridoma cell line into the peritoneal cavity of a suitable vertebrate host, such as a mouse. Monoclonal antibodies may then be harvested from the ascites fluid or the blood. Contaminants may be removed from the antibodies by conventional techniques, such as chromatography, gel filtration, precipitation, and extraction. The polypeptides of this invention may be used in the purification process in, for example, an affinity chromatography step.

[0390] Within certain embodiments, the use of antigen-binding fragments of antibodies may be preferred. Such fragments include Fab fragments, which may be prepared using standard techniques. Briefly, immunoglobulins may be purified from rabbit serum by affinity chromatography on Protein A bead columns (Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988) and digested by papain to yield Fab and Fc fragments. The Fab and Fc fragments may be separated by affinity chromatography on protein A bead columns.

[0391] Monoclonal antibodies of the present invention may be coupled to one or more therapeutic agents. Suitable agents in this regard include radionuclides, differentiation inducers, drugs, toxins, and derivatives thereof. Preferred radionuclides include ⁹⁰Y, ¹²³I, ¹²⁵I, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, and ²¹²Bi. Preferred drugs include methotrexate, and pyrimidine and purine analogs. Preferred differentiation inducers include phorbol esters and butyric acid. Preferred toxins include ricin, abrin, diptheria toxin, cholera toxin, gelonin, Pseudomonas exotoxin, Shigella toxin, and pokeweed antiviral protein.

[0392] A therapeutic agent may be coupled (e.g., covalently bonded) to a suitable monoclonal antibody either directly or indirectly (e.g., via a linker group). A direct reaction between an agent and an antibody is possible when each possesses a substituent capable of reacting with the other. For example, a nucleophilic group, such as an amino or sulfhydryl group, on one may be capable of reacting with a carbonyl-containing group, such as an anhydride or an acid halide, or with an alkyl group containing a good leaving group (e.g., a halide) on the other.

[0393] Alternatively, it may be desirable to couple a therapeutic agent and an antibody via a linker group. A linker group can function as a spacer to distance an antibody from an agent in order to avoid interference with binding capabilities. A linker group can also serve to increase the chemical reactivity of a substituent on an agent or an antibody, and thus increase the coupling efficiency. An increase in chemical reactivity may also facilitate the use of agents, or functional groups on agents, which otherwise would not be possible.

[0394] It will be evident to those skilled in the art that a variety of bifunctional or polyfunctional reagents, both homo- and hetero-functional (such as those described in the catalog of the Pierce Chemical Co., Rockford, Ill.), may be employed as the linker group. Coupling may be effected, for example, through amino groups, carboxyl groups, sulfhydryl groups or oxidized carbohydrate residues. There are numerous references describing such methodology, e.g., U.S. Pat. No. 4,671,958, to Rodwell et al.

[0395] Where a therapeutic agent is more potent when free from the antibody portion of the immunoconjugates of the present invention, it may be desirable to use a linker group which is cleavable during or upon internalization into a cell. A number of different cleavable linker groups have been described. The mechanisms for the intracellular release of an agent from these linker groups include cleavage by reduction of a disulfide bond (e.g., U.S. Pat. No. 4,489,710, to Spitler), by irradiation of a photolabile bond (e.g., U.S. Pat. No. 4,625,014, to Senter et al.), by hydrolysis of derivatized amino acid side chains (e.g., U.S. Pat. No. 4,638,045, to Kohn et al.), by serum complement-mediated hydrolysis (e.g., U.S. Pat. No. 4,671,958, to Rodwell et al.), and acid-catalyzed hydrolysis (e.g., U.S. Pat. No. 4,569,789, to Blattler et al.).

[0396] It may be desirable to couple more than one agent to an antibody. In one embodiment, multiple molecules of an agent are coupled to one antibody molecule. In another embodiment, more than one type of agent may be coupled to one antibody. Regardless of the particular embodiment, immunoconjugates with more than one agent may be prepared in a variety of ways. For example, more than one agent may be coupled directly to an antibody molecule, or linkers that provide multiple sites for attachment can be used. Alternatively, a carrier can be used.

[0397] A carrier may bear the agents in a variety of ways, including covalent bonding either directly or via a linker group. Suitable carriers include proteins such as albumins (e.g., U.S. Pat. No. 4,507,234, to Kato et al.), peptides and polysaccharides such as aminodextran (e.g., U.S. Pat. No. 4,699,784, to Shih et al.). A carrier may also bear an agent by noncovalent bonding or by encapsulation, such as within a liposome vesicle (e.g., U.S. Pat. Nos. 4,429,008 and 4,873,088). Carriers specific for radionuclide agents include radiohalogenated small molecules and chelating compounds. For example, U.S. Pat. No. 4,735,792 discloses representative radiohalogenated small molecules and their synthesis. A radionuclide chelate may be formed from chelating compounds that include those containing nitrogen and sulfur atoms as the donor atoms for binding the metal, or metal oxide, radionuclide. For example, U.S. Pat. No. 4,673,562, to Davison et al. discloses representative chelating compounds and their synthesis.

[0398] A variety of routes of administration for the antibodies and immunoconjugates may be used. Typically, administration will be intravenous, intramuscular, subcutaneous and the like. It will be evident that the precise dose of the antibody/immunoconjugate will vary depending upon the antibody used, the antigen density, and the rate of clearance of the antibody.

[0399] T Cells

[0400] Immunotherapeutic compositions may also, or alternatively, comprise T cells specific for HSV protein. Such cells may generally be prepared in vitro or ex vivo, using standard procedures. For example, T cells may be isolated from bone marrow, peripheral blood, or a fraction of bone marrow or peripheral blood of a patient, using a commercially available cell separation system, such as the Isolex™ System, available from Nexell Therapeutics, Inc. (Irvine, Calif.; see also U.S. Pat. No. 5,240,856; U.S. Pat. No. 5,215,926; WO 89/06280; WO 91/16116 and WO 92/07243). Alternatively, T cells may be derived from related or unrelated humans, non-human mammals, cell lines or cultures.

[0401] T cells may be stimulated with a HSV polypeptide, polynucleotide encoding a HSV polypeptide and/or an antigen presenting cell (APC) that expresses such a polypeptide. Such stimulation is performed under conditions and for a time sufficient to permit the generation of T cells that are specific for the polypeptide. In certain embodiments, HSV polypeptide or polynucleotide is present within a delivery vehicle, such as a microsphere, to facilitate the generation of specific T cells.

[0402] T cells are considered to be specific for a HSV polypeptide if the T cells specifically proliferate, secrete cytokines or kill target cells coated with the polypeptide or expressing a gene encoding the polypeptide. T cell specificity may be evaluated using any of a variety of standard techniques. For example, within a chromium release assay or proliferation assay, a stimulation index of more than two fold increase in lysis and/or proliferation, compared to negative controls, indicates T cell specificity. Such assays may be performed, for example, as described in Chen et al., Cancer Res. 54:1065-1070, 1994. Alternatively, detection of the proliferation of T cells may be accomplished by a variety of known techniques. For example, T cell proliferation can be detected by measuring an increased rate of DNA synthesis (e.g., by pulse-labeling cultures of T cells with tritiated thymidine and measuring the amount of tritiated thymidine incorporated into DNA). Contact with a HSV polypeptide (100 ng/ml-100 μg/ml, preferably 200 ng/ml-25 μg/ml) for 3-7 days should result in at least a two fold increase in proliferation of the T cells. Contact as described above for 2-3 hours should result in activation of the T cells, as measured using standard cytokine assays in which a two fold increase in the level of cytokine release (e.g., TNF or IFN-γ) is indicative of T cell activation (see Coligan et al., Current Protocols in Immunology, vol. 1, Wiley Interscience (Greene 1998)). T cells that have been activated in response to a HSV polypeptide, polynucleotide or polypeptide-expressing APC may be CD4⁺ and/or CD8⁺. HSV protein-specific T cells may be expanded using standard techniques. Within preferred embodiments, the T cells are derived from a patient, a related donor or an unrelated donor, and are administered to the patient following stimulation and expansion.

[0403] For therapeutic purposes, CD4⁺ or CD8⁺ T cells that proliferate in response to a HSV polypeptide, polynucleotide or APC can be expanded in number either in vitro or in vivo. Proliferation of such T cells in vitro may be accomplished in a variety of ways. For example, the T cells can be re-exposed to a HSV polypeptide, or a short peptide corresponding to an immunogenic portion of such a polypeptide, with or without the addition of T cell growth factors, such as interleukin-2, and/or stimulator cells that synthesize a HSV polypeptide. Alternatively, one or more T cells that proliferate in the presence of a HSV protein can be expanded in number by cloning. Methods for cloning cells are well known in the art, and include limiting dilution.

[0404] Pharmaceutical Compositions

[0405] In additional embodiments, the present invention concerns formulation of one or more of the polynucleotide, polypeptide, T-cell and/or antibody compositions disclosed herein in pharmaceutically-acceptable solutions for administration to a cell or an animal, either alone, or in combination with one or more other modalities of therapy.

[0406] It will also be understood that, if desired, the nucleic acid segment, RNA, DNA or PNA compositions that express a polypeptide as disclosed herein may be administered in combination with other agents as well, such as, e.g., other proteins or polypeptides or various pharmaceutically-active agents. In fact, there is virtually no limit to other components that may also be included, given that the additional agents do not cause a significant adverse effect upon contact with the target cells or host tissues. The compositions may thus be delivered along with various other agents as required in the particular instance. Such compositions may be purified from host cells or other biological sources, or alternatively may be chemically synthesized as described herein. Likewise, such compositions may further comprise substituted or derivatized RNA or DNA compositions.

[0407] Formulation of pharmaceutically-acceptable excipients and carrier solutions is well-known to those of skill in the art, as is the development of suitable dosing and treatment regimens for using the particular compositions described herein in a variety of treatment regimens, including e.g., oral, parenteral, intravenous, intranasal, and intramuscular administration and formulation.

[0408] 1. Oral Delivery

[0409] In certain applications, the pharmaceutical compositions disclosed herein may be delivered via oral administration to an animal. As such, these compositions may be formulated with an inert diluent or with an assimilable edible carrier, or they may be enclosed in hard- or soft-shell gelatin capsule, or they may be compressed into tablets, or they may be incorporated directly with the food of the diet.

[0410] The active compounds may even be incorporated with excipients and used in the form of ingestible tablets, buccal tables, troches, capsules, elixirs, suspensions, syrups, wafers, and the like (Mathiowitz et al., 1997; Hwang et al., 1998; U.S. Pat. No. 5,641,515; U.S. Pat. No. 5,580,579 and U.S. Pat. No. 5,792,451, each specifically incorporated herein by reference in its entirety). The tablets, troches, pills, capsules and the like may also contain the following: a binder, as gum tragacanth, acacia, cornstarch, or gelatin; excipients, such as dicalcium phosphate; a disintegrating agent, such as corn starch, potato starch, alginic acid and the like; a lubricant, such as magnesium stearate; and a sweetening agent, such as sucrose, lactose or saccharin may be added or a flavoring agent, such as peppermint, oil of wintergreen, or cherry flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellac, sugar, or both. A syrup of elixir may contain the active compound sucrose as a sweetening agent methyl and propylparabens as preservatives, a dye and flavoring, such as cherry or orange flavor. Of course, any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed. In addition, the active compounds may be incorporated into sustained-release preparation and formulations.

[0411] Typically, these formulations may contain at least about 0.1% of the active compound or more, although the percentage of the active ingredient(s) may, of course, be varied and may conveniently be between about 1 or 2% and about 60% or 70% or more of the weight or volume of the total formulation. Naturally, the amount of active compound(s) in each therapeutically useful composition may be prepared is such a way that a suitable dosage will be obtained in any given unit dose of the compound. Factors such as solubility, bioavailability, biological half-life, route of administration, product shelf life, as well as other pharmacological considerations will be contemplated by one skilled in the art of preparing such pharmaceutical formulations, and as such, a variety of dosages and treatment regimens may be desirable.

[0412] For oral administration the compositions of the present invention may alternatively be incorporated with one or more excipients in the form of a mouthwash, dentifrice, buccal tablet, oral spray, or sublingual orally-administered formulation. For example, a mouthwash may be prepared incorporating the active ingredient in the required amount in an appropriate solvent, such as a sodium borate solution (Dobell's Solution). Alternatively, the active ingredient may be incorporated into an oral solution such as one containing sodium borate, glycerin and potassium bicarbonate, or dispersed in a dentifrice, or added in a therapeutically-effective amount to a composition that may include water, binders, abrasives, flavoring agents, foaming agents, and humectants. Alternatively the compositions may be fashioned into a tablet or solution form that may be placed under the tongue or otherwise dissolved in the mouth.

[0413] 2. Injectable Delivery

[0414] In certain circumstances it will be desirable to deliver the pharmaceutical compositions disclosed herein parenterally, intravenously, intramuscularly, or even intraperitoneally as described in U.S. Pat. No. 5,543,158; U.S. Pat. No. 5,641,515 and U.S. Pat. No. 5,399,363 (each specifically incorporated herein by reference in its entirety). Solutions of the active compounds as free base or pharmacologically acceptable salts may be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0415] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions (U.S. Pat. No. 5,466,468, specifically incorporated herein by reference in its entirety). In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and/or vegetable oils. Proper fluidity may be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be facilitated by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0416] For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, a sterile aqueous medium that can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage may be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, “Remington's Pharmaceutical Sciences” 15th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. Moreover, for human administration, preparations should meet sterility, pyrogenicity, and the general safety and purity standards as required by FDA Office of Biologics standards.

[0417] Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0418] The compositions disclosed herein may be formulated in a neutral or salt form. Pharmaceutically-acceptable salts, include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms such as injectable solutions, drug-release capsules, and the like.

[0419] As used herein, “carrier” includes any and all solvents, dispersion media, vehicles, coatings, diluents, antibacterial and antifungal agents, isotonic and absorption delaying agents, buffers, carrier solutions, suspensions, colloids, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.

[0420] The phrase “pharmaceutically-acceptable” refers to molecular entities and compositions that do not produce an allergic or similar untoward reaction when administered to a human. The preparation of an aqueous composition that contains a protein as an active ingredient is well understood in the art. Typically, such compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid prior to injection can also be prepared. The preparation can also be emulsified.

[0421] 3. Nasal Delivery

[0422] In certain embodiments, the pharmaceutical compositions may be delivered by intranasal sprays, inhalation, and/or other aerosol delivery vehicles. Methods for delivering genes, nucleic acids, and peptide compositions directly to the lungs via nasal aerosol sprays has been described e.g., in U.S. Pat. No. 5,756,353 and U.S. Pat. No. 5,804,212 (each specifically incorporated herein by reference in its entirety). Likewise, the delivery of drugs using intranasal microparticle resins (Takenaga et al., 1998) and lysophosphatidyl-glycerol compounds (U.S. Pat. No. 5,725,871, specifically incorporated herein by reference in its entirety) are also well-known in the pharmaceutical arts. Likewise, transmucosal drug delivery in the form of a polytetrafluoroetheylene support matrix is described in U.S. Pat. No. 5,780,045 (specifically incorporated herein by reference in its entirety).

[0423] 4. Liposome-, Nanocapsule-, and Microparticle-Mediated Delivery

[0424] In certain embodiments, the inventors contemplate the use of liposomes, nanocapsules, microparticles, microspheres, lipid particles, vesicles, and the like, for the introduction of the compositions of the present invention into suitable host cells. In particular, the compositions of the present invention may be formulated for delivery either encapsulated in a lipid particle, a liposome, a vesicle, a nanosphere, or a nanoparticle or the like.

[0425] Such formulations may be preferred for the introduction of pharmaceutically-acceptable formulations of the nucleic acids or constructs disclosed herein. The formation and use of liposomes is generally known to those of skill in the art (see for example, Couvreur et al., 1977; Couvreur, 1988; Lasic, 1998; which describes the use of liposomes and nanocapsules in the targeted antibiotic therapy for intracellular bacterial infections and diseases). Recently, liposomes were developed with improved serum stability and circulation half-times (Gabizon and Papahadjopoulos, 1988; Allen and Choun, 1987; U.S. Pat. No. 5,741,516, specifically incorporated herein by reference in its entirety). Further, various methods of liposome and liposome like preparations as potential drug carriers have been reviewed (Takakura, 1998; Chandran et al., 1997; Margalit, 1995; U.S. Pat. No. 5,567,434; U.S. Pat. No. 5,552,157; U.S. Pat. No. 5,565,213; U.S. Pat. No. 5,738,868 and U.S. Pat. No. 5,795,587, each specifically incorporated herein by reference in its entirety).

[0426] Liposomes have been used successfully with a number of cell types that are normally resistant to transfection by other procedures including T cell suspensions, primary hepatocyte cultures and PC 12 cells (Renneisen et al., 1990; Muller et al., 1990). In addition, liposomes are free of the DNA length constraints that are typical of viral-based delivery systems. Liposomes have been used effectively to introduce genes, drugs (Heath and Martin, 1986; Heath et al., 1986; Balazsovits et al., 1989; Fresta and Puglisi, 1996), radiotherapeutic agents (Pikul et al., 1987), enzymes (Imaizumi et al., 1990a; Imaizumi et al., 1990b), viruses (Faller and Baltimore, 1984), transcription factors and allosteric effectors (Nicolau and Gersonde, 1979) into a variety of cultured cell lines and animals. In addition, several successful clinical trails examining the effectiveness of liposome-mediated drug delivery have been completed (Lopez-Berestein et al., 1985a; 1985b; Coune, 1988; Sculier et al., 1988). Furthermore, several studies suggest that the use of liposomes is not associated with autoimmune responses, toxicity or gonadal localization after systemic delivery (Mori and Fukatsu, 1992).

[0427] Liposomes are formed from phospholipids that are dispersed in an aqueous medium and spontaneously form multilamellar concentric bilayer vesicles (also termed multilamellar vesicles (MLVs). MLVs generally have diameters of from 25 nm to 4 μm. Sonication of MLVs results in the formation of small unilamellar vesicles (SUVs) with diameters in the range of 200 to 500 Å, containing an aqueous solution in the core.

[0428] Liposomes bear resemblance to cellular membranes and are contemplated for use in connection with the present invention as carriers for the peptide compositions. They are widely suitable as both water- and lipid-soluble substances can be entrapped, i.e., in the aqueous spaces and within the bilayer itself, respectively. It is possible that the drug-bearing liposomes may even be employed for site-specific delivery of active agents by selectively modifying the liposomal formulation.

[0429] In addition to the teachings of Couvreur et al. (1977; 1988), the following information may be utilized in generating liposomal formulations. Phospholipids can form a variety of structures other than liposomes when dispersed in water, depending on the molar ratio of lipid to water. At low ratios the liposome is the preferred structure. The physical characteristics of liposomes depend on pH, ionic strength and the presence of divalent cations. Liposomes can show low permeability to ionic and polar substances, but at elevated temperatures undergo a phase transition which markedly alters their permeability. The phase transition involves a change from a closely packed, ordered structure, known as the gel state, to a loosely packed, less-ordered structure, known as the fluid state. This occurs at a characteristic phase-transition temperature and results in an increase in permeability to ions, sugars and drugs.

[0430] In addition to temperature, exposure to proteins can alter the permeability of liposomes. Certain soluble proteins, such as cytochrome c, bind, deform and penetrate the bilayer, thereby causing changes in permeability. Cholesterol inhibits this penetration of proteins, apparently by packing the phospholipids more tightly. It is contemplated that the most useful liposome formations for antibiotic and inhibitor delivery will contain cholesterol.

[0431] The ability to trap solutes varies between different types of liposomes. For example, MLVs are moderately efficient at trapping solutes, but SUVs are extremely inefficient. SUVs offer the advantage of homogeneity and reproducibility in size distribution, however, and a compromise between size and trapping efficiency is offered by large unilamellar vesicles (LUVs). These are prepared by ether evaporation and are three to four times more efficient at solute entrapment than MLVs.

[0432] In addition to liposome characteristics, an important determinant in entrapping compounds is the physicochemical properties of the compound itself. Polar compounds are trapped in the aqueous spaces and nonpolar compounds bind to the lipid bilayer of the vesicle. Polar compounds are released through permeation or when the bilayer is broken, but nonpolar compounds remain affiliated with the bilayer unless it is disrupted by temperature or exposure to lipoproteins. Both types show maximum efflux rates at the phase transition temperature.

[0433] Liposomes interact with cells via four different mechanisms:

[0434] endocytosis by phagocytic cells of the reticuloendothelial system such as macrophages and neutrophils; adsorption to the cell surface, either by nonspecific weak hydrophobic or electrostatic forces, or by specific interactions with cell-surface components; fusion with the plasma cell membrane by insertion of the lipid bilayer of the liposome into the plasma membrane, with simultaneous release of liposomal contents into the cytoplasm; and by transfer of liposomal lipids to cellular or subcellular membranes, or vice versa, without any association of the liposome contents. It often is difficult to determine which mechanism is operative and more than one may operate at the same time.

[0435] The fate and disposition of intravenously injected liposomes depend on their physical properties, such as size, fluidity, and surface charge. They may persist in tissues for h or days, depending on their composition, and half lives in the blood range from min to several h. Larger liposomes, such as MLVs and LUVs, are taken up rapidly by phagocytic cells of the reticuloendothelial system, but physiology of the circulatory system restrains the exit of such large species at most sites. They can exit only in places where large openings or pores exist in the capillary endothelium, such as the sinusoids of the liver or spleen. Thus, these organs are the predominate site of uptake. On the other hand, SUVs show a broader tissue distribution but still are sequestered highly in the liver and spleen. In general, this in vivo behavior limits the potential targeting of liposomes to only those organs and tissues accessible to their large size. These include the blood, liver, spleen, bone marrow, and lymphoid organs.

[0436] Targeting is generally not a limitation in terms of the present invention. However, should specific targeting be desired, methods are available for this to be accomplished. Antibodies may be used to bind to the liposome surface and to direct the antibody and its drug contents to specific antigenic receptors located on a particular cell-type surface. Carbohydrate determinants (glycoprotein or glycolipid cell-surface components that play a role in cell-cell recognition, interaction and adhesion) may also be used as recognition sites as they have potential in directing liposomes to particular cell types. Mostly, it is contemplated that intravenous injection of liposomal preparations would be used, but other routes of administration are also conceivable.

[0437] Alternatively, the invention provides for pharmaceutically-acceptable nanocapsule formulations of the compositions of the present invention. Nanocapsules can generally entrap compounds in a stable and reproducible way (Henry-Michelland et al., 1987; Quintanar-Guerrero et al., 1998; Douglas et al., 1987). To avoid side effects due to intracellular polymeric overloading, such ultrafine particles (sized around 0.1 μm) should be designed using polymers able to be degraded in vivo. Biodegradable polyalkyl-cyanoacrylate nanoparticles that meet these requirements are contemplated for use in the present invention. Such particles may be are easily made, as described (Couvreur et al., 1980; 1988; zur Muhlen et al., 1998; Zambaux et al. 1998; Pinto-Alphandry et al., 1995 and U.S. Pat. No. 5,145,684, specifically incorporated herein by reference in its entirety).

[0438] Vaccines

[0439] In certain preferred embodiments of the present invention, vaccines are provided. The vaccines will generally comprise one or more pharmaceutical compositions, such as those discussed above, in combination with an immunostimulant. An immunostimulant may be any substance that enhances or potentiates an immune response (antibody and/or cell-mediated) to an exogenous antigen. Examples of immunostimulants include adjuvants, biodegradable microspheres (e.g., polylactic galactide) and liposomes (into which the compound is incorporated; see, e.g., Fullerton, U.S. Pat. No. 4,235,877). Vaccine preparation is generally described in, for example, M. F. Powell and M. J. Newman, eds., “Vaccine Design (the subunit and adjuvant approach),” Plenum Press (NY, 1995). Pharmaceutical compositions and vaccines within the scope of the present invention may also contain other compounds, which may be biologically active or inactive. For example, one or more immunogenic portions of other HSV antigens may be present, either incorporated into a fusion polypeptide or as a separate compound, within the composition or vaccine.

[0440] Illustrative vaccines may contain DNA encoding one or more of the polypeptides as described above, such that the polypeptide is generated in situ. As noted above, the DNA may be present within any of a variety of delivery systems known to those of ordinary skill in the art, including nucleic acid expression systems, bacteria and viral expression systems. Numerous gene delivery techniques are well known in the art, such as those described by Rolland, Crit. Rev. Therap. Drug Carrier Systems 15:143-198, 1998, and references cited therein. Appropriate nucleic acid expression systems contain the necessary DNA sequences for expression in the patient (such as a suitable promoter and terminating signal). Bacterial delivery systems involve the administration of a bacterium (such as Bacillus-Calmette-Guerrin) that expresses an immunogenic portion of the polypeptide on its cell surface or secretes such an epitope. In a preferred embodiment, the DNA may be introduced using a viral expression system (e.g., vaccinia or other pox virus, retrovirus, or adenovirus), which may involve the use of a non-pathogenic (defective), replication competent virus. Suitable systems are disclosed, for example, in Fisher-Hoch et al., Proc. Natl. Acad. Sci. USA 86:317-321,1989; Flexner et al., Ann. N. Y. Acad. Sci. 569:86-103, 1989; Flexner et al., Vaccine 8:17-21,1990; U.S. Pat. Nos. 4,603,112, 4,769,330, and 5,017,487; WO 89/01973; U.S. Pat. No. 4,777,127; GB 2,200,651; EP 0,345,242; WO 91/02805; Berkner, Biotechniques 6:616-627, 1988; Rosenfeld et al., Science 252:431-434,1991; Kolls et al., Proc. Natl. Acad. Sci. USA 91:215-219,1994; Kass-Eisler et al., Proc. Natl. Acad. Sci. USA 90:11498-11502,1993; Guzman et al., Circulation 88:2838-2848,1993; and Guzman et al., Cir. Res. 73:1202-1207,1993. Techniques for incorporating DNA into such expression systems are well known to those of ordinary skill in the art. The DNA may also be “naked,” as described, for example, in Ulmer et al., Science 259:1745-1749,1993 and reviewed by Cohen, Science 259:1691-1692,1993. The uptake of naked DNA may be increased by coating the DNA onto biodegradable beads, which are efficiently transported into the cells. It will be apparent that a vaccine may comprise both a polynucleotide and a polypeptide component. Such vaccines may provide for an enhanced immune response.

[0441] It will be apparent that a vaccine may contain pharmaceutically acceptable salts of the polynucleotides and polypeptides provided herein. Such salts may be prepared from pharmaceutically acceptable non-toxic bases, including organic bases (e.g., salts of primary, secondary and tertiary amines and basic amino acids) and inorganic bases (e.g., sodium, potassium, lithium, ammonium, calcium and magnesium salts).

[0442] While any suitable carrier known to those of ordinary skill in the art may be employed in the vaccine compositions of this invention, the type of carrier will vary depending on the mode of administration. Compositions of the present invention may be formulated for any appropriate manner of administration, including for example, topical, oral, nasal, intravenous, intracranial, intraperitoneal, subcutaneous or intramuscular administration. For parenteral administration, such as subcutaneous injection, the carrier preferably comprises water, saline, alcohol, a fat, a wax or a buffer. For oral administration, any of the above carriers or a solid carrier, such as mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, glucose, sucrose, and magnesium carbonate, may be employed. Biodegradable microspheres (e.g., polylactate polyglycolate) may also be employed as carriers for the pharmaceutical compositions of this invention. Suitable biodegradable microspheres are disclosed, for example, in U.S. Pat. Nos. 4,897,268; 5,075,109; 5,928,647; 5,811,128; 5,820,883; 5,853,763; 5,814,344 and 5,942,252. Modified hepatitis B core protein carrier systems are also suitable, such as those described in WO/99 40934, and references cited therein, all incorporated herein by reference. One may also employ a carrier comprising the particulate-protein complexes described in U.S. Pat. No. 5,928,647, which are capable of inducing a class I-restricted cytotoxic T lymphocyte responses in a host.

[0443] Such compositions may also comprise buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol, proteins, polypeptides or amino acids such as glycine, antioxidants, bacteriostats, chelating agents such as EDTA or glutathione, adjuvants (e.g., aluminum hydroxide), solutes that render the formulation isotonic, hypotonic or weakly hypertonic with the blood of a recipient, suspending agents, thickening agents and/or preservatives. Alternatively, compositions of the present invention may be formulated as a lyophilizate. Compounds may also be encapsulated within liposomes using well known technology.

[0444] Any of a variety of immunostimulants may be employed in the vaccines of this invention. For example, an adjuvant may be included. Most adjuvants contain a substance designed to protect the antigen from rapid catabolism, such as aluminum hydroxide or mineral oil, and a stimulator of immune responses, such as lipid A, Bortadella pertussis or Mycobacterium tuberculosis derived proteins. Suitable adjuvants are commercially available as, for example, Freund's Incomplete Adjuvant and Complete Adjuvant (Difco Laboratories, Detroit, Mich.); Merck Adjuvant 65 (Merck and Company, Inc., Rahway, N.J.); AS-2 (SmithKline Beecham, Philadelphia, Pa.); aluminum salts such as aluminum hydroxide gel (alum) or aluminum phosphate; salts of calcium, iron or zinc; an insoluble suspension of acylated tyrosine; acylated sugars; cationically or anionically derivatized polysaccharides; polyphosphazenes; biodegradable microspheres; monophosphoryl lipid A and quil A. Cytokines, such as GM-CSF or interleukin-2, -7, or -12, may also be used as adjuvants.

[0445] Within the vaccines provided herein, the adjuvant composition is preferably designed to induce an immune response predominantly of the Th1 type. High levels of Th1-type cytokines (e.g., IFN-γ, TNFα, IL-2 and IL-12) tend to favor the induction of cell mediated immune responses to an administered antigen. In contrast, high levels of Th2-type cytokines (e.g., IL-4, IL-5, IL-6 and IL-10) tend to favor the induction of humoral immune responses. Following application of a vaccine as provided herein, a patient will support an immune response that includes Th1- and Th2-type responses. Within a preferred embodiment, in which a response is predominantly Th1-type, the level of Th1-type cytokines will increase to a greater extent than the level of Th2-type cytokines. The levels of these cytokines may be readily assessed using standard assays. For a review of the families of cytokines, see Mosmann and Coffman, Ann. Rev. Immunol. 7:145-173, 1989.

[0446] Preferred adjuvants for use in eliciting a predominantly Th1-type response include, for example, a combination of monophosphoryl lipid A, preferably 3-de-O-acylated monophosphoryl lipid A (3D-MPL), together with an aluminum salt. MPL adjuvants are available from Corixa Corporation (Seattle, Wash.; see U.S. Pat. Nos. 4,436,727; 4,877,611; 4,866,034 and 4,912,094). CpG-containing oligonucleotides (in which the CpG dinucleotide is unmethylated) also induce a predominantly Th1 response. Such oligonucleotides are well known and are described, for example, in WO 96/02555, WO 99/33488 and U.S. Pat. Nos. 6,008,200 and 5,856,462. Immunostimulatory DNA sequences are also described, for example, by Sato et al., Science 273:352,1996. Another preferred adjuvant is a saponin, preferably QS21 (Aquila Biopharmaceuticals Inc., Framingham, Mass.), which may be used alone or in combination with other adjuvants. For example, an enhanced system involves the combination of a monophosphoryl lipid A and saponin derivative, such as the combination of QS21 and 3D-MPL as described in WO 94/00153, or a less reactogenic composition where the QS21 is quenched with cholesterol, as described in WO 96/33739. Other preferred formulations comprise an oil-in-water emulsion and tocopherol. A particularly potent adjuvant formulation involving QS21, 3D-MPL and tocopherol in an oil-in-water emulsion is described in WO 95/17210.

[0447] Other preferred adjuvants include Montanide ISA 720 (Seppic, France), SAF (Chiron, California, United States), ISCOMS (CSL), MF-59 (Chiron), the SBAS series of adjuvants (e.g., SBAS-2 or SBAS-4, available from SmithKline Beecham, Rixensart, Belgium), Detox (Corixa, Hamilton, Mont.), RC-529 (Corixa, Hamilton, Mont.) and other aminoalkyl glucosaminide 4-phosphates (AGPs), such as those described in pending U.S. patent application Ser. Nos. 08/853,826 and 09/074,720, the disclosures of which are incorporated herein by reference in their entireties. Other preferred adjuvants comprise polyoxyethylene ethers, such as those described in WO 99/52549A1.

[0448] Any vaccine provided herein may be prepared using well known methods that result in a combination of antigen, immune response enhancer and a suitable carrier or excipient. The compositions described herein may be administered as part of a sustained release formulation (i.e., a formulation such as a capsule, sponge or gel (composed of polysaccharides, for example) that effects a slow release of compound following administration). Such formulations may generally be prepared using well known technology (see, e.g., Coombes et al., Vaccine 14:1429-1438, 1996) and administered by, for example, oral, rectal or subcutaneous implantation, or by implantation at the desired target site. Sustained-release formulations may contain a polypeptide, polynucleotide or antibody dispersed in a carrier matrix and/or contained within a reservoir surrounded by a rate controlling membrane.

[0449] Carriers for use within such formulations are biocompatible, and may also be biodegradable; preferably the formulation provides a relatively constant level of active component release. Such carriers include microparticles of poly(lactide-co-glycolide), polyacrylate, latex, starch, cellulose, dextran and the like. Other delayed-release carriers include supramolecular biovectors, which comprise a non-liquid hydrophilic core (e.g., a cross-linked polysaccharide or oligosaccharide) and, optionally, an external layer comprising an amphiphilic compound, such as a phospholipid (see, e.g., U.S. Pat. No. 5,151,254 and PCT applications WO 94/20078, WO/94/23701 and WO 96/06638). The amount of active compound contained within a sustained release formulation depends upon the site of implantation, the rate and expected duration of release and the nature of the condition to be treated or prevented.

[0450] Any of a variety of delivery vehicles may be employed within pharmaceutical compositions and vaccines to facilitate production of an antigen-specific immune response that targets HSV-infected cells. Delivery vehicles include antigen presenting cells (APCs), such as dendritic cells, macrophages, B cells, monocytes and other cells that may be engineered to be efficient APCs. Such cells may, but need not, be genetically modified to increase the capacity for presenting the antigen, to improve activation and/or maintenance of the T cell response, to have anti-HSV effects per se and/or to be immunologically compatible with the receiver (i.e., matched HLA haplotype). APCs may generally be isolated from any of a variety of biological fluids and organs and may be autologous, allogeneic, syngeneic or xenogeneic cells.

[0451] Certain preferred embodiments of the present invention use dendritic cells or progenitors thereof as antigen-presenting cells. Dendritic cells are highly potent APCs (Banchereau and Steinman, Nature 392:245-251,1998) and have been shown to be effective as a physiological adjuvant for eliciting prophylactic or therapeutic immunity (see Timmerman and Levy, Ann. Rev. Med. 50:507-529, 1999). In general, dendritic cells may be identified based on their typical shape (stellate in situ, with marked cytoplasmic processes (dendrites) visible in vitro), their ability to take up, process and present antigens with high efficiency and their ability to activate naive T cell responses. Dendritic cells may, of course, be engineered to express specific cell-surface receptors or ligands that are not commonly found on dendritic cells in vivo or ex vivo, and such modified dendritic cells are contemplated by the present invention. As an alternative to dendritic cells, secreted vesicles antigen-loaded dendritic cells (called exosomes) may be used within a vaccine (see Zitvogel et al., Nature Med. 4:594-600, 1998).

[0452] Dendritic cells and progenitors may be obtained from peripheral blood, bone marrow, lymph nodes, spleen, skin, umbilical cord blood or any other suitable tissue or fluid. For example, dendritic cells may be differentiated ex vivo by adding a combination of cytokines such as GM-CSF, IL-4, IL-13 and/or TNFα to cultures of monocytes harvested from peripheral blood. Alternatively, CD34 positive cells harvested from peripheral blood, umbilical cord blood or bone marrow may be differentiated into dendritic cells by adding to the culture medium combinations of GM-CSF, IL-3, TNFα, CD40 ligand, LPS, flt3 ligand and/or other compound(s) that induce differentiation, maturation and proliferation of dendritic cells.

[0453] Dendritic cells are conveniently categorized as “immature” and “mature” cells, which allows a simple way to discriminate between two well characterized phenotypes. However, this nomenclature should not be construed to exclude all possible intermediate stages of differentiation. Immature dendritic cells are characterized as APC with a high capacity for antigen uptake and processing, which correlates with the high expression of Fcγ receptor and mannose receptor. The mature phenotype is typically characterized by a lower expression of these markers, but a high expression of cell surface molecules responsible for T cell activation such as class I and class II MHC, adhesion molecules (e.g., CD54 and CD11) and costimulatory molecules (e.g., CD40, CD8O, CD86 and 4-1 BB).

[0454] APCs may generally be transfected with a polynucleotide encoding a HSV protein (or portion or other variant thereof) such that the HSV polypeptide, or an immunogenic portion thereof, is expressed on the cell surface. Such transfection may take place ex vivo, and a composition or vaccine comprising such transfected cells may then be used for therapeutic purposes, as described herein. Alternatively, a gene delivery vehicle that targets a dendritic or other antigen presenting cell may be administered to a patient, resulting in transfection that occurs in vivo. In vivo and ex vivo transfection of dendritic cells, for example, may generally be performed using any methods known in the art, such as those described in WO 97/24447, or the gene gun approach described by Mahvi et al., Immunology and cell Biology 75:456-460,1997. Antigen loading of dendritic cells may be achieved by incubating dendritic cells or progenitor cells with the HSV polypeptide, DNA (naked or within a plasmid vector) or RNA; or with antigen-expressing recombinant bacterium or viruses (e.g., vaccinia, fowlpox, adenovirus or lentivirus vectors). Prior to loading, the polypeptide may be covalently conjugated to an immunological partner that provides T cell help (e.g., a carrier molecule). Alternatively, a dendritic cell may be pulsed with a non-conjugated immunological partner, separately or in the presence of the polypeptide.

[0455] Vaccines and pharmaceutical compositions may be presented in unit-dose or multi-dose containers, such as sealed ampoules or vials. Such containers are preferably hermetically sealed to preserve sterility of the formulation until use. In general, formulations may be stored as suspensions, solutions or emulsions in oily or aqueous vehicles. Alternatively, a vaccine or pharmaceutical composition may be stored in a freeze-dried condition requiring only the addition of a sterile liquid carrier immediately prior to use.

[0456] Immunotherapeutic Applications

[0457] In further aspects of the present invention, the compositions described herein may be used for immunotherapy of HSV infections. Within such methods, pharmaceutical compositions and vaccines are typically administered to a patient. As used herein, a “patient” refers to any warm-blooded animal, preferably a human. The above pharmaceutical compositions and vaccines may be used to prophylactically prevent or ameliorate the extent of infection by HSV or to treat a patient already infected with HSV. Administration may be by any suitable method, including administration by intravenous, intraperitoneal, intramuscular, subcutaneous, intranasal, intradermal, anal, vaginal, topical, and oral routes.

[0458] Within certain embodiments, immunotherapy may be active immunotherapy, in which treatment relies on the in vivo stimulation of the endogenous host immune system to react against HSV infection with the administration of immune response-modifying agents (such as polypeptides and polynucleotides as provided herein).

[0459] Within other embodiments, immunotherapy may be passive immunotherapy, in which treatment involves the delivery of agents with established HSV-immune reactivity (such as effector cells or antibodies) that can directly or indirectly mediate therapeutic effects and does not necessarily depend on an intact host immune system. Examples of effector cells include T cells as discussed above, T lymphocytes (such as CD8⁺ cytotoxic T lymphocytes and CD4⁺ T-helper lymphocytes), killer cells (such as Natural Killer cells and lymphokine-activated killer cells), B cells and antigen-presenting cells (such as dendritic cells and macrophages) expressing a polypeptide provided herein. T cell receptors and antibody receptors specific for the polypeptides recited herein may be cloned, expressed and transferred into other vectors or effector cells for adoptive immunotherapy. The polypeptides provided herein may also be used to generate antibodies or anti-idiotypic antibodies (as described above and in U.S. Pat. No. 4,918,164) for passive immunotherapy.

[0460] Effector cells may generally be obtained in sufficient quantities for adoptive immunotherapy by growth in vitro, as described herein. Culture conditions for expanding single antigen-specific effector cells to several billion in number with retention of antigen recognition in vivo are well known in the art. Such in vitro culture conditions typically use intermittent stimulation with antigen, often in the presence of cytokines (such as IL-2) and non-dividing feeder cells. As noted above, immunoreactive polypeptides as provided herein may be used to rapidly expand antigen-specific T cell cultures in order to generate a sufficient number of cells for immunotherapy. In particular, antigen-presenting cells, such as dendritic, macrophage, monocyte, fibroblast and/or B cells, may be pulsed with immunoreactive polypeptides or transfected with one or more polynucleotides using standard techniques well known in the art. For example, antigen-presenting cells can be transfected with a polynucleotide having a promoter appropriate for increasing expression in a recombinant virus or other expression system. Cultured effector cells for use in therapy must be able to grow and distribute widely, and to survive long term in vivo. Studies have shown that cultured effector cells can be induced to grow in vivo and to survive long term in substantial numbers by repeated stimulation with antigen supplemented with IL-2 (see, for example, Cheever et al., Immunological Reviews 157:177,1997).

[0461] Alternatively, a vector expressing a polypeptide recited herein may be introduced into antigen presenting cells taken from a patient and clonally propagated ex vivo for transplant back into the same patient. Transfected cells may be reintroduced into the patient using any means known in the art, preferably in sterile form by intravenous, intracavitary or intraperitoneal.

[0462] Routes and frequency of administration of the therapeutic compositions described herein, as well as dosage, will vary from individual to individual, but may be readily established using standard techniques. In one embodiment, between 1 and about 10 doses may be administered over a 52 week period. In another embodiment, about 6 doses are administered, at intervals of about 1 month, and booster vaccinations are typically be given periodically thereafter. Alternate protocols may be appropriate for individual patients.

[0463] A suitable dose is an amount of a compound that, when administered as described above, is capable of promoting an anti-HSV immune response, and is preferably at least 10-50% above the basal (i.e., untreated) level. Such response can be monitored, for example, by measuring the anti-HSV antibodies in a patient. Such vaccines should also be capable of causing an immune response that leads to an improved clinical outcome (e.g., more frequent remissions, complete or partial or longer disease-free survival) in vaccinated patients as compared to non-vaccinated patients. In general, for pharmaceutical compositions and vaccines comprising one or more polypeptides, the amount of each polypeptide present in a dose ranges from about 25 μg to 5 mg per kg of host. Suitable dose sizes will vary with the size of the patient, but will typically range from about 0.1 mL to about 5 mL.

[0464] In general, an appropriate dosage and treatment regimen provides the active compound(s) in an amount sufficient to provide therapeutic and/or prophylactic benefit. Such a response can be monitored by establishing an improved clinical outcome (e.g., more frequent remissions, complete or partial, or longer disease-free survival) in treated patients as compared to non-treated patients. Increases in preexisting immune responses to a HSV protein may correlate with an improved clinical outcome. Such immune responses may generally be evaluated using standard proliferation, cytotoxicity or cytokine assays, which may be performed using samples obtained from a patient before and after treatment.

[0465] HSV Detection and Diagnosis

[0466] In general, HSV may be detected in a patient based on the presence of one or more HSV proteins and/or polynucleotides encoding such proteins in a biological sample (for example, blood, sera, sputum urine and/or other appropriate tissue) obtained from the patient. In other words, such proteins may be used as markers to indicate the presence or absence of HSV in a patient. The binding agents provided herein generally permit detection of the level of antigen that binds to the agent in the biological sample. Polynucleotide primers and probes may be used to detect the level of mRNA encoding a HSV protein, which is also indicative of the presence or absence of HSV infection.

[0467] There are a variety of assay formats known to those of ordinary skill in the art for using a binding agent to detect polypeptide markers in a sample. See, e.g., Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988. In general, the presence or absence of HSV in a patient may be determined by contacting a biological sample obtained from a patient with a binding agent and detecting in the sample a level of polypeptide that binds to the binding agent.

[0468] In a preferred embodiment, the assay involves the use of binding agent immobilized on a solid support to bind to and remove the polypeptide from the remainder of the sample. The bound polypeptide may then be detected using a detection reagent that contains a reporter group and specifically binds to the binding agent/polypeptide complex. Such detection reagents may comprise, for example, a binding agent that specifically binds to the polypeptide or an antibody or other agent that specifically binds to the binding agent, such as an anti-immunoglobulin, protein G, protein A or a lectin. Alternatively, a competitive assay may be utilized, in which a polypeptide is labeled with a reporter group and allowed to bind to the immobilized binding agent after incubation of the binding agent with the sample. The extent to which components of the sample inhibit the binding of the labeled polypeptide to the binding agent is indicative of the reactivity of the sample with the immobilized binding agent. Suitable polypeptides for use within such assays include full length HSV proteins and portions thereof to which the binding agent binds, as described above.

[0469] The solid support may be any material known to those of ordinary skill in the art to which the protein may be attached. For example, the solid support may be a test well in a microtiter plate or a nitrocellulose or other suitable membrane. Alternatively, the support may be a bead or disc, such as glass, fiberglass, latex or a plastic material such as polystyrene or polyvinylchloride. The support may also be a magnetic particle or a fiber optic sensor, such as those disclosed, for example, in U.S. Pat. No. 5,359,681. The binding agent may be immobilized on the solid support using a variety of techniques known to those of skill in the art, which are amply described in the patent and scientific literature. In the context of the present invention, the term “immobilization” refers to both noncovalent association, such as adsorption, and covalent attachment (which may be a direct linkage between the agent and functional groups on the support or may be a linkage by way of a cross-linking agent). Immobilization by adsorption to a well in a microtiter plate or to a membrane is preferred. In such cases, adsorption may be achieved by contacting the binding agent, in a suitable buffer, with the solid support for a suitable amount of time. The contact time varies with temperature, but is typically between about 1 hour and about 1 day. In general, contacting a well of a plastic microtiter plate (such as polystyrene or polyvinylchloride) with an amount of binding agent ranging from about 10 ng to about 10 μg, and preferably about 100 ng to about 1 μg, is sufficient to immobilize an adequate amount of binding agent.

[0470] Covalent attachment of binding agent to a solid support may generally be achieved by first reacting the support with a bifunctional reagent that will react with both the support and a functional group, such as a hydroxyl or amino group, on the binding agent. For example, the binding agent may be covalently attached to supports having an appropriate polymer coating using benzoquinone or by condensation of an aldehyde group on the support with an amine and an active hydrogen on the binding partner (see, e.g., Pierce Immunotechnology Catalog and Handbook, 1991, at A12-A13).

[0471] In certain embodiments, the assay is a two-antibody sandwich assay. This assay may be performed by first contacting an antibody that has been immobilized on a solid support, commonly the well of a microtiter plate, with the sample, such that polypeptides within the sample are allowed to bind to the immobilized antibody. Unbound sample is then removed from the immobilized polypeptide-antibody complexes and a detection reagent (preferably a second antibody capable of binding to a different site on the polypeptide) containing a reporter group is added. The amount of detection reagent that remains bound to the solid support is then determined using a method appropriate for the specific reporter group.

[0472] More specifically, once the antibody is immobilized on the support as described above, the remaining protein binding sites on the support are typically blocked. Any suitable blocking agent known to those of ordinary skill in the art, such as bovine serum albumin or Tween ₂₀™ (Sigma Chemical Co., St. Louis, Mo.). The immobilized antibody is then incubated with the sample, and polypeptide is allowed to bind to the antibody. The sample may be diluted with a suitable diluent, such as phosphate-buffered saline (PBS) prior to incubation. In general, an appropriate contact time (i.e., incubation time) is a period of time that is sufficient to detect the presence of polypeptide within a sample obtained from an individual with an HSV infection. Preferably, the contact time is sufficient to achieve a level of binding that is at least about 95% of that achieved at equilibrium between bound and unbound polypeptide. Those of ordinary skill in the art will recognize that the time necessary to achieve equilibrium may be readily determined by assaying the level of binding that occurs over a period of time. At room temperature, an incubation time of about 30 minutes is generally sufficient.

[0473] Unbound sample may then be removed by washing the solid support with an appropriate buffer, such as PBS containing 0.1% Tween 20™. The second antibody, which contains a reporter group, may then be added to the solid support. Preferred reporter groups include those groups recited above.

[0474] The detection reagent is then incubated with the immobilized antibody-polypeptide complex for an amount of time sufficient to detect the bound polypeptide. An appropriate amount of time may generally be determined by assaying the level of binding that occurs over a period of time. Unbound detection reagent is then removed and bound detection reagent is detected using the reporter group. The method employed for detecting the reporter group depends upon the nature of the reporter group. For radioactive groups, scintillation counting or autoradiographic methods are generally appropriate. Spectroscopic methods may be used to detect dyes, luminescent groups and fluorescent groups. Biotin may be detected using avidin, coupled to a different reporter group (commonly a radioactive or fluorescent group or an enzyme). Enzyme reporter groups may generally be detected by the addition of substrate (generally for a specific period of time), followed by spectroscopic or other analysis of the reaction products.

[0475] To determine the presence or absence of HSV, the signal detected from the reporter group that remains bound to the solid support is generally compared to a signal that corresponds to a predetermined cut-off value. In one embodiment, the cut-off value for the detection of HSV is the average mean signal obtained when the immobilized antibody is incubated with samples from patients without HSV. In an alternate embodiment, the cut-off value is determined using a Receiver Operator Curve, according to the method of Sackett et al., Clinical Epidemiology: A Basic Science for Clinical Medicine, Little Brown and Co., 1985, p. 106-7. Briefly, in this embodiment, the cut-off value may be determined from a plot of pairs of true positive rates (i.e., sensitivity) and false positive rates (100%-specificity) that correspond to each possible cut-off value for the diagnostic test result. The cut-off value on the plot that is the closest to the upper left-hand corner (i.e., the value that encloses the largest area) is the most accurate cut-off value, and a sample generating a signal that is higher than the cut-off value determined by this method may be considered positive. Alternatively, the cut-off value may be shifted to the left along the plot, to minimize the false positive rate, or to the right, to minimize the false negative rate. In general, a sample generating a signal that is higher than the cut-off value determined by this method is considered positive.

[0476] In a related embodiment, the assay is performed in a flow-through or strip test format, wherein the binding agent is immobilized on a membrane, such as nitrocellulose. In the flow-through test, polypeptides within the sample bind to the immobilized binding agent as the sample passes through the membrane. A second, labeled binding agent then binds to the binding agent-polypeptide complex as a solution containing the second binding agent flows through the membrane. The detection of bound second binding agent may then be performed as described above. In the strip test format, one end of the membrane to which binding agent is bound is immersed in a solution containing the sample. The sample migrates along the membrane through a region containing second binding agent and to the area of immobilized binding agent. Concentration of second binding agent at the area of immobilized antibody indicates the presence of HSV. Typically, the concentration of second binding agent at that site generates a pattern, such as a line, that can be read visually. The absence of such a pattern indicates a negative result. In general, the amount of binding agent immobilized on the membrane is selected to generate a visually discernible pattern when the biological sample contains a level of polypeptide that would be sufficient to generate a positive signal in the two-antibody sandwich assay, in the format discussed above. Preferred binding agents for use in such assays are antibodies and antigen-binding fragments thereof. Preferably, the amount of antibody immobilized on the membrane ranges from about 25 ng to about 1 μg, and more preferably from about 50 ng to about 500 ng. Such tests can typically be performed with a very small amount of biological sample.

[0477] Of course, numerous other assay protocols exist that are suitable for use with the HSV proteins or binding agents of the present invention. The above descriptions are intended to be exemplary only. For example, it will be apparent to those of ordinary skill in the art that the above protocols may be readily modified to use HSV polypeptides to detect antibodies that bind to such polypeptides in a biological sample. The detection of such protein-specific antibodies can allow for the identification of HSV infection.

[0478] HSV infection may also, or alternatively, be detected based on the presence of T cells that specifically react with a HSV protein in a biological sample. Within certain methods, a biological sample comprising CD4⁺ and/or CD8⁺ T cells isolated from a patient is incubated with a HSV polypeptide, a polynucleotide encoding such a polypeptide and/or an APC that expresses at least an immunogenic portion of such a polypeptide, and the presence or absence of specific activation of the T cells is detected. Suitable biological samples include, but are not limited to, isolated T cells. For example, T cells may be isolated from a patient by routine techniques (such as by Ficoll/Hypaque density gradient centrifugation of peripheral blood lymphocytes). T cells may be incubated in vitro for about 2-9 days (typically about 4 days) at 37° C. with polypeptide (e.g., 5-25 μg/ml). It may be desirable to incubate another aliquot of a T cell sample in the absence of HSV polypeptide to serve as a control. For CD4⁺ T cells, activation is preferably detected by evaluating proliferation of the T cells. For CD8⁺ T cells, activation is preferably detected by evaluating cytolytic activity. A level of proliferation that is at least two fold greater and/or a level of cytolytic activity that is at least 20% greater than in disease-free patients indicates the presence of HSV in the patient.

[0479] As noted above, HSV infection may also, or alternatively, be detected based on the level of mRNA encoding a HSV protein in a biological sample. For example, at least two oligonucleotide primers may be employed in a polymerase chain reaction (PCR) based assay to amplify a portion of a HSV cDNA derived from a biological sample, wherein at least one of the oligonucleotide primers is specific for (i.e., hybridizes to) a polynucleotide encoding the HSV protein. The amplified cDNA is then separated and detected using techniques well known in the art, such as gel electrophoresis. Similarly, oligonucleotide probes that specifically hybridize to a polynucleotide encoding a HSV protein may be used in a hybridization assay to detect the presence of polynucleotide encoding the HSV protein in a biological sample.

[0480] To permit hybridization under assay conditions, oligonucleotide primers and probes should comprise an oligonucleotide sequence that has at least about 60%, preferably at least about 75% and more preferably at least about 90%, identity to a portion of a polynucleotide encoding a HSV protein that is at least 10 nucleotides, and preferably at least 20 nucleotides, in length. Preferably, oligonucleotide primers and/or probes hybridize to a polynucleotide encoding a polypeptide described herein under moderately stringent conditions, as defined above. Oligonucleotide primers and/or probes which may be usefully employed in the diagnostic methods described herein preferably are at least 10-40 nucleotides in length. In a preferred embodiment, the oligonucleotide primers comprise at least 10 contiguous nucleotides, more preferably at least 15 contiguous nucleotides, of a DNA molecule having a sequence disclosed herein. Techniques for both PCR based assays and hybridization assays are well known in the art (see, for example, Mullis et al., Cold Spring Harbor Symp. Quant. Biol., 51:263, 1987; Erlich ed., PCR Technology, Stockton Press, NY, 1989).

[0481] One preferred assay employs RT-PCR, in which PCR is applied in conjunction with reverse transcription. Typically, RNA is extracted from a biological sample, such as biopsy tissue, and is reverse transcribed to produce cDNA molecules. PCR amplification using at least one specific primer generates a cDNA molecule, which may be separated and visualized using, for example, gel electrophoresis. Amplification may be performed on biological samples taken from a test patient and from an individual who is not infected with HSV. The amplification reaction may be performed on several dilutions of cDNA, for example spanning two orders of magnitude.

[0482] As noted above, to improve sensitivity, multiple HSV protein markers may be assayed within a given sample. It will be apparent that binding agents specific for different HSV polypeptides may be combined within a single assay. Further, multiple primers or probes may be used concurrently. The selection of HSV protein markers may be based on routine experiments to determine combinations that results in optimal sensitivity. In addition, or alternatively, assays for HSV proteins provided herein may be combined with assays for other known HSV antigens.

[0483] The present invention further provides kits for use within any of the above diagnostic and/or therapeutic methods. Such kits typically comprise two or more components necessary for performing a diagnostic and/or therapeutic assay and will further comprise instructions for the use of said kit. Components may be compounds, reagents, containers and/or equipment. For example, one container within a diagnostic kit may contain a monoclonal antibody or fragment thereof that specifically binds to a HSV protein. Such antibodies or fragments may be provided attached to a support material, as described above. One or more additional containers may enclose elements, such as reagents or buffers, to be used in the assay. Such kits may also, or alternatively, contain a detection reagent as described above that contains a reporter group suitable for direct or indirect detection of antibody binding.

[0484] Alternatively, a kit may be designed to detect the level of mRNA encoding a HSV protein in a biological sample. Such kits generally comprise at least one oligonucleotide probe or primer, as described above, that hybridizes to a polynucleotide encoding a HSV protein. Such an oligonucleotide may be used, for example, within a PCR or hybridization assay. Additional components that may be present within such kits include a second oligonucleotide and/or a diagnostic reagent or container to facilitate the detection of a polynucleotide encoding a HSV protein.

[0485] The following Examples are offered by way of illustration and not by way of limitation.

EXAMPLE 1 Identification of HSV-2 Antigens

[0486] The following examples are presented to illustrate certain embodiments of the present invention and to assist one of ordinary skill in making and using the same. The examples are not intended in any way to otherwise limit the scope of the invention.

[0487] Source of HSV-2 Positive Donors:

[0488] Lymphocytes were obtained from two types of donors: Group A) seropositive donors with unknown clinical status, and Group B) seropositive donors with well characterized clinical status (viral shedding and ano-genital lesion recurrences).

[0489] Group A: Blood samples (50 ml) were obtained from 13 potential donors. No information regarding clinical history of HSV-2 infection was requested. The blood was screened for serum antibody against HSV-1 and HSV-2 by Western blot. PBMCs were also screened for specific proliferative T cell responses to HSV-1 and HSV-2 lysate antigens (ABI; Columbia, Md.). Three donors (AD104, AD116, and AD120) were positive for HSV-2 serum antibody and their PBMCs specifically proliferated in response to HSV-2 antigen. Leukopheresis PBMC were collected from these donors and cryopreserved in liquid nitrogen.

[0490] Group B: Ano-genital lesion biopisies were obtained from donors DK21318 and JR5032. Lesion biopsy lymphocytes were expanded in vitro with IL-2 and PHA in the presence of 50 uM acyclovir and subsequently cryopreserved in liquid nitrogen. Typically 5×10⁶ to 5×10⁷ lymphocytes are obtained after two weeks. Autologous PBMC were also collected from the blood of DK2318 and JR5032 and cryopreserved in liquid nitrogen.

[0491] Generation of CD4⁺ T Cell Lines:

[0492] Cryopreserved PBMCs or lesion-biopsy lymphocytes were thawed and stimulated in vitro with 1 ug/ml HSV-2 antigen (ABI) in RPMI 1640+10% human serum+10 ng/ml IL-7. Irradiated autologous PBMC were added as antigen presenting cells for the lesion biopsy lymphocytes only. Recombinant IL-2 (1 ng/ml) was added on days 1 and 4. The cells were harvested, washed, and replated in fresh medium containing IL-2 and IL-7 on day 7. Recombinant IL-2 was again added on day 10. The T cells were harvested, washed, and restimulated in vitro with HSV-2 antigen plus irradiated autologous PBMCin the same manner on day 14 of culture. The T cell lines were cryopreserved at 1×10⁷cells/vial in liquid nitrogen on day 11-12 of the secondary stimulation. After thawing, the cryopreserved T cells retained the ability to specifically proliferate to HSV-2 antigen in vitro. These T cells were subsequently used to screen HSV-2 gene-fragment expression cloning libraries prepared in E. coli, as described below.

[0493] Preparation of HSV-2 (333) DNA:

[0494] HSV-2 strain 333 virus was grown in Vero cells cultured in roller bottles in 200 ml/bottle of Medium 199 (Gibco)+5% FCS. Vero cells are transformed African green monkey fibroblast-like cells that were obtained from ATCC (Cat. #CCL-81). Near-confluence Vero cells (10 roller bottles) were infected with HSV-2 strain 333 virus at an MOI of 0.01 in 50 ml/bottle of Medium 199+1% FCS. Cells and medium were harvested from the roller bottles and the cells pelleted. The supernatant was saved on ice and the cell pellets were resuspended in fresh Medium 199+1% FCS and lysed by 6 cycles of freezing/thawing. The cell debris in the lysates was pelleted and the supernatant pooled with the saved culture supernatant. Virus was pelleted from the pooled supernatants by ultracentrifugation (12,000g, 2 hours, 4° C.) and resuspended in 2 ml of fresh Medium 199+1% FCS. The virus was further purified on a 5-15% linear Ficoll gradient by ultracentrifugation (19,000 g, 2 hours, 4° C.) as previously described (Chapter 10:Herpes simplex virus vectors of Molecular Virology: A Practical Approach (1993); Authors: F. J. Rixon and J. McClaughlan, Editors: A. J. Davison and R. M. Elliott; Publisher: Oxford University Press, Inc, New York, N.Y.). The HSV-2 virus-containing band was extracted from the gradient, diluted 10-fold with Medium 199, and the virus pelleted by ultracentrifugation at 19,000 g for 4 hours at 4° C. The virus pellet was recovered and resuspended in 10 ml of Tris/EDTA (TE) buffer. Intact virions were treated with DNAse and RNAse to remove cellular DNA and RNA. The enzymes were then inactivated by addition of EDTA and incubation at 65° C. DNA was prepared from the gradient-purified virus by lysis of the viral particles with SDS in the presence of EDTA, followed by phenol/chlorform extraction to purify the genomic viral DNA. HSV-2 DNA was precipitated with EtOH and the DNA pellet was dried and resuspended in 1 ml of Tris/EDTA buffer. The concentration and purity of the DNA was determined by reading the OD 260 and OD 280 on a UV spectrophotometer. Genomic DNA prepared in this manner was used for construction of an HSV-2 genomic fragment expression library in E. coli.

[0495] Construction of HSV-2 DNA Fragment Libraries in the pET17b Vector:

[0496] The HSV2-I library was constructed as follows. DNA fragments were generated by sonicating genomic HSV-2 DNA for 4 seconds at 15% output with a Fisher “60 SonicDismembrator” (Fisher). The sonicated DNA was then precipitated, pelleted, and resuspended in 11 uL TE buffer. The approximate size of the DNA fragments was measured by agarose gel electropheresis of 1 uL of the fragmented HSV-2 genomic DNA vs. 1.5 ug unsonicated material. The average size of the DNA fragments was determined to be approx. 500 bp when visualized after ethidium bromide staining of the gel. Incomplete DNA fragment ends were filled in (blunted) using T4 DNA polymerase. EcoR1 adapters were then ligated to the blunt ends of the DNA fragments using T4 DNA ligase. The DNA was then kinased using T4 Polynucleotide Kinase, purified using a manually loaded column of S-400-HR Sephacryl (Sigma) and ligated into the pET17b expression vector. The HSV2-l library was constructed in a similar fashion. The average size of inserts in this library was determined to be approximately 1000 bp.

[0497] Generation of the HSV-2 Fragment Expression Library in E. coli.

[0498] The HSV2-1 library was transformed into E. coli for preparation of glycerol stocks and testing of HSV-2 DNA insert representation. The DNA was transformed into ElectroMAX DH10B E. coli (Gibco) in order to prepare a large quantity of HSV-2/pET17b library DNA. Transformed bacteria were grown up on 3 LB/Ampicillin plates (approx. 750 CFU/plate), a small subset of colonies were picked for sequencing of DNA inserts, and the remaining bacteria from each plate collected as a pool for preparation of plasmid DNA. These pools were named HSV-2 Pools 9, 10 and 11. Glycerol stocks of a portion of these bacterial pools were stored at −80° C. Plasmids were purified from the remainder of the pools. Equal quantities of plasmid DNA from each of the 3 pools was combined to make a single pool of plasmid DNA. The tranformation efficiency of the pooled DNA was empirically determined using JM109(DE3) E. coli bacteria. JM109(DE3) bacteria were then transformed with an amount of the final pool of library DNA that was expected to yield 15 colony-forming units (CFU) per plate. The transformed bacteria were then plated on 100 LB/amp plates. Twenty CFU (on average) were actually observed on each of the 100 plates; therefore the pool size of this HSV-2 library was about 20 clones/pool. The bacterial colonies were collected as a pool from each plate in approximately 800 ul/plate of LB+20% glycerol. Each pool was distributed equally (200 ul/well) among four 96-well U-bottom plates and these “master stock” plates were stored at −80° C. The size of this HSV-2 gene-fragment library (hereafter referred to as HSV21) was therefore 96 pools of 20 clones/pool. Plasmid DNA was prepared from 20 randomly picked colonies and the inserts sequenced. Approximately 15% (3/20) contained HSV-2 DNA as insert, 80% (16/20) contained non-HSV-2 DNA (E. coli or Vero cell DNA), and 5% ({fraction (1/20)}) contained no insert DNA. The HSV2-II DNA library was transformed into E. coli and random colonies analyzed in a similar manner. Relevant differences in the construction of library HSV2-II included the transformation of the HSV-2/pET17b ligation product into NovaBlue (Novagen) chemically competent E. coli instead of using electroporation for preparation of a larger quantity of plasmid for pooling and transformation into JM109(DE3) bacteria for empirical evaluation. Additionally, plasmid DNA was prepared from 10 pools averaging 160 colonies/plate. These 10 plasmid pools were combined in an equivalent fashion (normalized based on spectrophotometer readings) into one pool for transformation into JM109(DE3) as per previously, yielding an average of 20 colonies(clones)/plate for harvesting into glycerol stock pools as before. Approximately 25% contained HSV-2 DNA as insert, with the remaining 75% containing E. coli DNA as insert.

[0499] Induction of the HSV-2 Fragment Expression Library for Screening with Human CD4+ T Cells.

[0500] One of the master HSV21 library 96-well plates was thawed at room temperature. An aliquot (20 uL) was transferred from each well to a new 96 well plate containing 180 uL/well of LB medium +ampicillin. The bacteria were grown up overnight and then 40 ul transferred into two new 96-well plates containing 160 uL 2× YT medium+ampicillin. The bacteria were grown for 1 hr.15 min at 37° C. Protein expression was then induced by addition of IPTG to 200 mM. The bacteria were cultured for an additional 3 hrs. One of these plates was used to obtain spectrophotometer readings to normalize bacterial numbers/well. The second, normalized plate was used for screening with CD4+ T cells after pelleting the bacteria (approx. 2×10⁷/well) and removing the supernatants. The HSV2-II library was grown and induced in a similar fashion.

[0501] Preparation of Autologous Dendritic APC's:

[0502] Dendritic cells (DCs) were generated by culture of plastic-adherent donor cells (derived from 1×10⁸ PBMC) in 6 well plates (Costar 3506) in RPMI 1640+10% of a 1:1 mix of FCS:HS+10 ng/ml GM-CSF +10 ng/ml IL-4 at 37° C. Non-adherent DCs were collected from plates on day 6 of culture and irradiated with 3300 Rads. The DCs were then plated at 1×10⁴/well in flat-bottom 96-well plates (Costar 3596) and cultured overnight at 37° C. The following day, the DCs were pulsed with the induced HSV2-I or HSV2-II library pools by resuspending the bacterial pellets in 200 ul RPMI 1640+10% FCS without antibiotics and transferring 10 ul/well to the wells containing the DCs in 190 ul of the same medium without antibiotics. The DCs and bacteria were co-cultured for 90 minutes at 37° C. The DCs were then washed and resuspended in 100 ul/well RPMI 1640+10% HS+L-glut. +50 ug/ml gentamicin antibiotic.

[0503] Preparation of Responder T Cells:

[0504] Cryopreserved CD4+T cell lines were thawed 5 days before use and cultured at 37° C. in RPMI 1640+10% HS+1 ng/ml IL-2+10 ng/ml IL-7. After 2 days, the medium was replaced with fresh medium without IL-2 and IL-7.

[0505] Primary Screening of the HSV2 Libraries:

[0506] The T cells were resuspended in fresh RPMI 1640+10% HS and added at 2×10⁴/well to the plates containing the E. coli-pulsed autologous DC's. After 3 days, 100 ul/well of supernatant was removed and transferred to new 96 well plates. Half of the supernatant was subsequently tested for IFN-gamma content by ELISA and the remainder was stored at −20° C. The T cells were then pulsed with 1 uCi/well of [³H]-Thymidine (Amersham/Pharmacia; Piscataway, N.J.) for about 8 hours at 37° C. The 3H-pulsed cells were then harvested onto UniFilter GF/C plates (Packard; Downers Grove, Ill.) and the CPM of [3H]-incorporated subsequently measured using a scintillation counter (Top-Count; Packard). ELISA assays were performed on cell supernatants following a standard cytokine-capture ELISA protocol for human IFN-g.

[0507] From the HSV2-I library screening with T cells from Dl 04, wells HSV2I_H10 and HSV2I_H12, for which both CPM and IFN-g levels were significantly above background, were scored as positive.

[0508] Breakdown of Positive HSV2I Library Pools:

[0509] The positive wells (HSV2I_H10 and HSV2I_H12) from the initial CD4+ T cell screening experiment were grown up again from the master glycerol stock plate. Forty-eight sub-clones from each pool were randomly picked, grown up and IPTG-induced as described previously. The subclones were screened against the AD104 CD4+ T cell line as described above. A clone (HSV2I_H12A12) from the HSV2I_H12 pool breakdown scored positive. This positive result was verified in a second AD104 CD4+ T cell assay.

[0510] Identification of UL39 as a CD4+ T Cell Antigen:

[0511] The positive clone (HSV2I_H12A12) was subcloned and 10 clones picked for restriction digest analysis with EcoRI NB#675 pg. 34. All 10 clones contained DNA insert of the same size (approximately 900 bp in length). Three of these clones (HSV2I_H12A12_(—)1, 7, and 8) were chosen for sequencing and all contained identical insert sequences at both the 5′ and 3′ ends of the inserts. The DNA sequence of the insert is set forth in SEQ ID NO:1, and contains an open reading frame set forth in SEQ ID NO:2. The insert sequence was compared to the complete genomic sequence of HSV-2 strain HG52 (NCBI site, Accession #Z86099) and the sequence was determined have a high degree of homology to UL39 (a.k.a. ICP6), the large subunit (140 kD) of the HSV ribonucleotide reductase, the sequence of which is set forth in SEQ ID NO:3. The insert sequence set forth in SEQ ID NO: 1 spans nucleotides 876-1690 of the UL39 open reading frame (3,432 bp) and encodes the amino acid sequence set forth in SEQ ID NO:2, which has a high degree of homology to amino acids 292-563 of UL39 (full length =1143 aa).

[0512] Identification of US8A. US3/US4. UL15, UL18, UL27 and UL46 as CD4+ T Cell Antigens:

[0513] In a manner essentially identical to that described above for the identification of UL39 as a T cell antigen, an additional HSV2 gene fragment expression cloning library, referred to as HSV2-II, was prepared, expressed in E. coli, and screened with donor T cells.

[0514] Screening the HSV2-II library with T cells from donor AD116 identified the clone HSV2II_US8AfragD6.B_B11_T7Trc.seq, determined to have an insert sequence set forth in SEQ ID NO:4, encoding open reading frames having amino acid sequences set forth in SEQ ID NO:5 and 6, with the sequence of SEQ ID NO:5 having a high degree of homology with the HSV2 US8A protein, the sequence of which is set forth in SEQ ID NO:7.

[0515] In addition, screening the HSV2-II library with T cells from donor AD104 identified the following clone inserts:

[0516] SEQ ID NO:8, corresponding to clone HSV2II_US3/US4 fragF10B3_T7Trc.seq, containing a potential open reading frame having an amino acid sequence set forth in SEQ ID NO: 10;

[0517] SEQ ID NO:9, corresponding to clone HSV2II_US3/US4 fragF10B3_T7P.seq, containing an open reading frame having an amino acid sequence set forth in SEQ ID NO: 11, sharing a high degree of homology with the HSV-2 US3 protein (SEQ ID NO: 12);

[0518] SEQ ID NO:13, corresponding to clone HSV2II_UL46fragF11F5_T7Trc.seq, containing an open reading frame having an amino acid sequence set forth in SEQ ID NO: 14, sharing a high degree of homology with the HSV-2 UL46 protein (SEQ ID NO: 15);

[0519] SEQ ID NO:16, corresponding to clone HSV2II_UL27frag-H2C7_T7Trc.seq, containing an open reading frame having an amino acid sequence set forth in SEQ ID NO:17, sharing a high degree of homology with the HSV-2 UL27 protein (SEQ ID NO:18);

[0520] SEQ ID NO:19, corresponding to clone HSV2II_UL18fragF10A1_rc.seq, containing open reading frames having amino acid sequences set forth in SEQ ID NO:20, 21 and 22, with SEQ ID NO:22 sharing a high degree of homology with the HSV-2 UL18 protein (SEQ ID NO: 23); and

[0521] SEQ ID NO:24, corresponding to clone HSV2I_UL15fragF10A12_rc.seq, containing an open reading frame having an amino acid sequence set forth in SEQ ID NO: 25, sharing a high degree of homology with the HSV-2 UL15 protein (SEQ ID NO: 26).

EXAMPLE 2 Identification of HSV-2 Antigens

[0522] CD4⁺ T cells from AD104 were found to recognize inserts from clones HSV2II_UL46fragF11F5_T7Trc.seq (SEQ ID NO: 13) and HSV2II_UL18frgaF10A1_rc.seq (SEQ ID NO: 19) as described in detail in Example 1. The sequences from these clones share a high degree of homology to the HSV2-I genes, UL46 (SEQ ID NO: 15) and UL18 (SEQ ID NO:23), respectively. Therefore to further characterize the epitopes recognized by these T cells, overlapping 15-mer peptides were made across the clone insert fragments of UL18 and UL46. Peptide recognition by AD104's CD4+ T cells was tested in a 48 hour IFN-g ELISPOT assay. ELISPOTS were performed by adding 1×10⁴ autologous EBV-transformed B cells (LCL) or DCs per well in 96 well ELISPOT plates. 2×10⁴ AD104 CD4+ T cells from AD104's line were added per well with 5 μg/ml of the HSV2 peptides. AD104 CD4+ T cells recognized peptides 20 and 21 (SEQ ID NO: 32 and 33) of UL18, and peptides 1, 4, 9, 10, and 20 of UL46 (SEQ ID NO: 27-31).

EXAMPLE 3 Identification of HSV-2 Antigens

[0523] CD4+ T cell lines were generated from DK2318 and JR5032 lesion-biopsy. The CD4+ lymphocytes were stimulated twice in vitro on irradiated autologous PBMC and HSV2 antigen as described in example 1. The lines were tested for their antigen specificity as described in example 1 and cryopreserved. The CD4+ T cell lines were screened against the HSV2-II expression-cloning library generated in Example 1.

[0524] DK2318 was shown to react with clones C12 and G10. Clone C12 was determined to have an insert sequence set forth in SEQ ID NO:36. This insert was found to have sequence homology with fragments of 2 HSV-11 genes, nucleotides 723-1311 of UL23 and nucleotides 1-852 of UL22. These sequences correspond to amino acids 241-376 of UL23 as set forth in SEQ ID NO:40 and amino acids 1-284 as set forth in SEQ ID NO:41. The DNA sequence of SEQ ID NO:36 was searched against public databases including Genbank and shown to have a high degree of sequence homology to the HSV2 genes UL23 and UL22 set forth in SEQ ID NO:37 and 38 respectively. The protein sequences encoded by SEQ ID NO:37 and 38 are set forth in SEQ ID NO:39 and 45. Clone G10 was determined to have an insert sequence which is set forth in SEQ ID NO:48, encoding open reading frames having an amino acid sequence set forth in SEQ ID NO:50, with the sequence of SEQ ID NO:48 having a high degree of sequence homology with HSV2 UL37, the sequence of which is set forth in SEQ ID NO:49, encoding open reading frames having the amino acid sequences set forth in SEQ ID NO:51. DK2318's CD4+ T cell line was screened against overlapping 15 mers covering the UL23 protein. DK2318's CD4 line was shown to react against three UL23 specific peptides (SEQ ID NO:41-43) suggesting that UL23 is a target.

[0525] The CD4+ T cell line generated from JR5032 was found to react with clone E9 which contained an insert sequence set forth in SEQ ID NO: 34, encoding open reading frames having amino acid sequences set forth in SEQ ID NO: 46, with SEQ ID NO: 34 having a high degree of sequence homology with HSV2 RL2 (also referred to as ICP0), the sequence of which is set forth in SEQ ID NO:35, encoding an open reading frame having the amino acid sequences set forth in SEQ ID NO:47.

EXAMPLE 4 Characterization of CD4 Clones F11F5 And G10A9

[0526] Examples 2 and 3 describe the generation of CD4 T cell lines from donors AD104 and DK2313 which were screened against cDNA libraries generated using the HSV-2333 strain. AD104 was found to react against the clone HSV2II_UL46fragF11F5. This insert was partially sequenced with the sequence being disclosed in SEQ ID NO:13. Full length sequencing of the insert revealed that it encoded a fragment of UL46 which was derived from the HSV-2 333 strain. The DNA and amino acid sequences from this insert are disclosed in SEQ ID NO:52 and 54, respectively.

[0527] DK2312 was found to react against the clone G10. This insert was partially sequenced and the sequence was disclosed in SEQ ID NO:48. Full length sequencing revealed that it encoded a fragment of UL37 which was derived from the HSV-2333 strain. The DNA and amino acid sequences from this insert are disclosed in SEQ ID NO:53 and 55, respectively.

EXAMPLE 5 Identification of CD8-Specific Immunoreactive Peptides Derived from HSV-2

[0528] Peripheral blood mononuclear cells were obtained from the normal donors AD104, AD116, AD120, and D477. These donors were HLA typed using low-resolution DNA-typing methodology and the results are presented in Table 2. TABLE 2 DONOR AD104 AD116 AD120 D477 HLA-A 24, 33 0206, 24 0211, 3303 0201, 2501 HLA-B 45, 58 0702, 35 1505, 4403 1501, 5101 HLA-C 01, 0302 0702, 1203 0303, 0706 0304, 12

[0529] In order to determine which epitopes of HSV-2 were immunoreactive, synthetic peptides were synthesized. These peptides were 15 amino acids in length overlapping by 11 amino acids. The peptides were synthesized across the following regions of the following HSV-2 genes: UL15 (aa 600-734), UL18 (aa 1-110), UL23 (aa 241-376), UL46 (aa 617-722), US3 (aa125-276), and US8A (aa 83-146).

[0530] CD8⁺ T cells were purified from the PBMC of each of the donors described above using negative selection. The purified CD8+ T cells were then tested for their reactivity against the HSV-2 specific peptides. Co-cultures containing 2×10⁵ CD8⁺ T cells, 1×10⁴ autologous dendritic cells and 10 μg/ml of a peptide pool (on average containing 10 peptides/pool) were established in 96 well ELISPOT plates that had been pre-coated with anti-human IFN-γ antibody (1D1K: mAbTech). After 24 hours, the ELISPOT plates were developed using a standard protocol well known to one of skill in the art. The number of spots per well were then counted using an automated video microscopy ELISPOT plate reader. CD8+ T cells from donors demonstrating a positive response against a peptide pool were then subsequently tested against the individual peptides in that pool in a second ELISPOT assay. The results of peptide reactivity are presented in Table 3. TABLE 3 Peptide # Donor HSV-2 Gene (amino acid numbering) SEQ ID NO AD104 US3 #33 (262-276) 63 AD116 UL15 #23 (688-702) 56 #30 (716-730) 57 UL23  #7 (265-279) 58 UL46  #2 (621-635) 59  #8 (645-659) 60  #9 (649-663) 61 #11 (657-671) 62 US8A  #5 (99-113)  64 AD120 UL46 Peptides: #1-12 — D477 UL18 Peptides: #1-12 — UL23 Peptides: #1-20 — UL46 Peptides: #1-12 —

EXAMPLE 6 Identification of HSV-2 Antigens using CD4+ T Cell Cloning

[0531] This Example describes the generation of CD4⁺ T cell clones from two donors. Donor JH is an HSV-2 seropositive donor who experiences infrequent recurrences of genital lesions and sheds virus infrequently, as determined by virus culture and PCR on daily swabs). HH is an HSV-2 exposed, but HSV-2 seronegative donor.

[0532] CD4⁺ T cell clones for JH were generated by stimulating the donor's peripheral blood mononuclear cells (PBMC) for 14 days with UV-inactivated HSV-2, strain 333. Following two weeks of stimulation, the cells were cloned into 96 well plates using limiting dilution, and stimulated non-selectively using a monoclonal antibody against CD3. Following 2 weeks of expansion, the clones were tested for their reactivity against UV-inactivated HSV-2, gB2 protein, gD2 protein and UL50. Clones 5 and 34 recognized gB2, clone 30 recognized gD2, and clone 11 recognized UL50.

[0533] Clones 39 and 47 were used for expression cloning. Antigen presenting cells (APC) used for both the expansion of the T cells and for the expression cloning were derived from HLA-matched normal donors. The clones were screened against two HSV-2 specific libraries, HSV2-II and HSV2-III.

[0534] Clone 39 was found to specifically recognize a partial sequence from UL39 presented by the HSV2-III library pools 1F4, 1G2, 2C4, and 3G11. The full length DNA sequence of UL39 is disclosed in SEQ ID NO:65, with the corresponding protein sequence disclosed in SEQ ID NO:3. The specific DNA sequence from pools 1F4, 1G2, and 3G11 that Clone 39 reacted against were identical. The inserts were found to be 875 bp in length and the DNA sequence is disclosed in SEQ ID NO:66, with the corresponding amino acid sequence disclosed in SEQ ID NO:74. The insert from pool 2C4 was found to be 800 bp in length, the DNA sequence of which is disclosed in SEQ ID NO:67, with the corresponding amino acid sequence disclosed in SEQ ID NO:75.

[0535] Clone 47 was found to specifically recognize a partial sequence from ICP0 (RL2) presented by the HSV-2III library pools 2B2, 3A1, 3F12, 3H6, and 4B2. The full length DNA sequence of ICP0 was disclosed in SEQ ID NO:35, with the corresponding protein sequence disclosed in SEQ ID NO:47. The sequence inserts from pools 3H6, 3F12, and 4B2 were found to be identical, with an insert size of 1100 bp. The DNA sequence corresponding to the 5′ end of this sequence is disclosed in SEQ ID NO:68, with the 3′ end disclosed in SEQ ID NO:69. The insert from pool 3A1 was found to be 1000 bp in length, with the 5′ portion of the DNA sequence disclosed in SEQ ID NO:70 and the 3′ end of the insert disclosed in SEQ ID NO:71. The insert from pool 2B2 was found to be 1300 bp in length. The DNA sequence corresponding to the 5′ end of the insert is disclosed in SEQ ID NO:72, with the 3′ end of the sequence disclosed in SEQ ID NO:73.

[0536] CD4⁺ T cell clones for HH were generated by stimulating the donors peripheral blood mononuclear cells (PBMC) for 14 days with UV-inactivated HSV-2, strain 333. Following two weeks of stimulation, the cells were cloned into 96 well plates using limiting dilution, and stimulated non-selectively using PHA. The clones were screened for their ability to proliferate in response to both HSV-1 and HSV-2 proteins. Clones 6,18, 20, 22, 24, 27, 28, 29, 41, and 45 were all found to react strongly against HSV-1, however only clones 6, 18, 20, 22, and 24 were found to respond strongly to HSV-2. Therefore, clones 6, 18, 20, 22, and 24 were selected for expression cloning use. APC from an HLA-matched donor were used for in vitro expansion of the clones and for expression cloning. The clones were screened against two HSV-2 specific libraries, HSV2-II and HSV2-III (see Example 1 for details of libraries).

[0537] Clone 22 was found to recognize UL46 presented by the HSV2-II library, pools F7 and F11, in addition to pool 4E8 that was derived from the HSV2-III library.

EXAMPLE 7 Generation of a UL19 Expressing Vaccinia Virus

[0538] The UL19 gene was cloned into the Western Reserve Strain of Vaccinia Virus. This viral vector allows expression of UL19 in any cell infected with the vaccinia virus, or additionally, the vaccinia virus can be used to immunize humans or animals to generate immune responses against UL19.

[0539] In order to generate the vaccinia virus expressing UL19, the UL19 open reading frame (ORF), the sequence of which is disclosed in SEQ ID NO:76, was cloned from HSV-2 and inserted into the vaccinia virus shuttle plasmid, pSC11 (the DNA sequence of which is disclosed in SEQ ID NO:77). CV-1 cells transfected with the shuttle vector, pSC11/UL19, were co-infected with the wild-type Western Reserve Vaccinia Virus. In some cells, the shuttle plasmid underwent homologous recombination with the vaccinia virus, inserting the UL19 gene into the thymidine kinase location. These recombinant virions were isolated by plaque purification of 5-Bromo-deoxyuridine (BrdU) resistant virus that expressed Beta-galactosidase. The purified virus can then be used to infect cells to express the UL19 protein.

EXAMPLE 8 Generation of a UL47 Expressing Vaccinia Virus

[0540] The UL47 gene was cloned into the Western Reserve Strain of Vaccinia Virus. This viral vector allows expression of UL47 in any cell infected with the vaccinia virus, or additionally, the vaccinia virus can be used to immunize humans or animals to generate immune responses against UL47.

[0541] In order to generate the vaccinia virus expressing UL47, the UL47 ORF, the sequence of which is disclosed in SEQ ID NO:78, was cloned from HSV-2 and inserted into the vaccinia virus shuttle plasmid, pSC11 (the DNA sequence of which is disclosed in SEQ ID NO:77). CV-1 cells transfected with the shuttle vector, pSC11/UL47, were co-infected with the wild-type Western Reserve Vaccinia Virus. In some cells, the shuttle plasmid underwent homologous recombination with the vaccinia virus, inserting the UL47 gene into the thymidine kinase location. These recombinant virions were isolated by plaque purification of 5-Bromo-deoxyuridine (BrdU) resistant virus that expressed Beta-galactosidase. The purified virus can then be used to infect cells to express the UL47 protein.

EXAMPLE 9 Generation of a UL50 Expressing Vaccinia Virus

[0542] The UL50 gene was cloned into the Western Reserve Strain of Vaccinia Virus. This viral vector allows expression of UL50 in any cell infected with the vaccinia virus, or additionally, the vaccinia virus can be used to immunize humans or animals to generate immune responses against UL50.

[0543] In order to generate the vaccinia virus expressing UL50, the UL50 ORF, the sequence of which is disclosed in SEQ ID NO:79, was cloned from HSV-2 and inserted into the vaccinia virus shuttle plasmid, pSC11 (the DNA sequence of which is disclosed in SEQ ID NO:77). CV-1 cells transfected with the shuttle vector, pSC11/UL50, were co-infected with the wild-type Western Reserve Vaccinia Virus. In some cells, the shuttle plasmid underwent homologous recombination with the vaccinia virus, inserting the UL50 gene into the thymidine kinase location. These recombinant virions were isolated by plaque purification of 5-Bromo-deoxyuridine (BrdU) resistant virus that expressed Beta-galactosidase. The purified virus can then be used to infect cells to express the UL50 protein.

EXAMPLE 10 Generation of a UL49 Expressing Vaccinia Virus

[0544] To facilitate intracellular degradation and Class I presentation of the Herpes Simplex Virus gene, UL49 (the DNA sequence of which is disclosed in SEQ ID NO:81), a fusion of the human Ubiquitin gene (the DNA sequence of which is disclosed in SEQ ID NO:80) and UL49 was constructed with the Ubiquitin gene located 5′ of the UL49 gene. The last amino acid of the Ubiquitin ORF was mutated from glycine to alanine to prevent co-translational cleavage of the fusion protein. After assembly of the fusion by PCR, it was cloned into the vaccinia virus shuttle vector, pSC11 (the DNA sequence of which is disclosed in SEQ ID NO:77). CV-1 cells transfected with the shuttle vector, pSC11/ubiquitin-UL49, were co-infected with the wild type Western Reserve Vaccinia Virus. In some cells the shuttle plasmid underwent homologous recombination with the virus inserting the ubiquitin-UL49 gene into the thymidine kinase location. These recombinant virions were isolated by plaque purification of 5-Bromo-deoxyuridine (BrdU) resistant virus that expresses Beta-galactosidase. The purified virus can then be used to infect cells to express the UL49 protein.

[0545] The cells engineered to express UL49 are used to assay for specific immune responses to UL49 protein. This vaccinia virus vector can also be used as a vaccine in humans to generate preventative or therapeutic responses against HSV-2.

EXAMPLE 11 Expression of Herpes Simplex Virus Antigens in E. coli

[0546] This example describes the expression of recombinant HSV antigens using an E. coli expression system combined with an N-terminal histadine tag.

[0547] Expression of HSV UL21 in E. coli:

[0548] The HSV UL21 coding region (the DNA sequence of which is disclosed in SEQ ID NO:85) was PCR amplified with the following primers: PDM-602 (SEQ ID NO:98) 5′gagctcagctatgccaccacc3′ PDM-603 (SEQ ID NO:99) 5′cggcgaattcattagtagaggcggtggaaaaag3′

[0549] The PCR was Performed with the Following Reaction Components:

[0550] 10 μl 10× Pfu buffer

[0551] 1 μl 10 mM dNTPs

[0552] 2 μl 10 μM of each primer

[0553] 83 μl of sterile water

[0554] 1.5 μl Pfu DNA polymerase (Stratagene, La Jolla, Calif.)

[0555] 50 ng DNA

[0556] PCR Amplification was Performed Using the Following Reaction Conditions:

[0557] 96° C. for 2 minutes, followed by 40 cycles of:

[0558] 96° C. for 20 seconds;

[0559] 60° C. for 15 seconds; and

[0560] 72° C. for 2 minutes, followed by a final extension step of:

[0561] 72° C. for 4 minutes.

[0562] The PCR product was digested with EcoRI and cloned into pPDM His that had been cut with Eco72I and EcoRI. The amino acid sequence for the UL21-His construct was confirmed, and is disclosed in SEQ ID NO:91. The construct was then transformed into BLR pLys and BLR Codon Plus RP cells.

[0563] Expression of HSV UL39 in E. coli:

[0564] The HSV UL39 coding region (the DNA sequence of which is disclosed in SEQ ID NO:89) was PCR amplified from clone pET17b with the following primers: (SEQ ID NO:100) PDM-466 5′cacgccgccgcaccccaggcggac 3′ (SEQ ID NO:101) PDM-467 5′cggcgaattcattagtagaggcggtggaaaaag 3′

[0565] The PCR was Performed with the Following Reaction Components:

[0566] 10 μl 10× Pfu buffer

[0567] 1 μl 10 mM dNTPs

[0568] 2 μl 10 μM of each primer

[0569] 83 μl of sterile water

[0570] 1.5 μl Pfu DNA polymerase (Stratagene, La Jolla, Calif.)

[0571] 50 ng DNA

[0572] PCR Amplification was Performed Using the Following Reaction Conditions:

[0573] 96° C. for 2 minutes, followed by 40 cycles of:

[0574] 96° C. for 20 seconds;

[0575] 66° C. for 15 seconds; and

[0576] 72° C. for 2 minutes, followed by a final extension step of:

[0577] 72° C. for 4 minutes.

[0578] The PCR product was digested with EcoRI and cloned into pPDM His that had been cut with Eco72I and EcoRI. The amino acid sequence for the UL39-His construct was confirmed, and is disclosed in SEQ ID NO:90. The construct was then transformed into BLR pLys and BLR Codon Plus RP cells.

[0579] Expression of HSV UL49 in E. coli:

[0580] The HSV UL49 coding region (the DNA sequence of which is disclosed in SEQ ID NO:83) was PCR amplified from clone pET17b with the following primers: (SEQ ID NO:102) PDM-466: 5′cacacctctcgccgctccgtcaagtc 3′ (SEQ ID NO:103) PDM-467: 5′cataagaattcactactcgagggggcggggacg 3′

[0581] The PCR was Performed with the Following Reaction Components:

[0582] 10 μl 10× Pfu buffer

[0583] 10 μl 10× PCRx enhancer solution

[0584] 3 μl 10 mM dNTPs

[0585] 3 μl 50 mM mgSO₄

[0586] 2 μl 10 μM of each primer

[0587] 68 μl of sterile water

[0588] 1.0 μl Pfx polymerase (Gibco)

[0589] 50 ng DNA

[0590] PCR Amplification was Performed Using the Following Reaction Conditions:

[0591] 96° C. for 2 minutes, followed by 40 cycles of:

[0592] 96° C. for 20 seconds;

[0593] 67° C. for 15 seconds; and

[0594] 72° C. for 2 minutes, followed by a final extension step of:

[0595] 72° C. for 4 minutes.

[0596] The PCR product was digested with EcoRI and cloned into pPDM His that had been cut with Eco 72I and EcoRI. The amino acid sequence for the UL49-His construct was confirmed, and is disclosed in SEQ ID NO:97. The construct was then transformed into BLR pLys and BLR Codon Plus RP cells.

[0597] Expression of HSV UL50 in E. coli:

[0598] The HSV UL50 coding region (the DNA sequence of which is disclosed in SEQ ID NO:82) was PCR amplified from clone pET17b with the following primers: (SEQ ID NO:104) PDM-458: 5′cacagtcagtgggggcccagggcgatcc 3′ (SEQ ID NO:105) PDM-459: 5′cctagaattcactagatgccagtggagccaaaccc 3′

[0599] The PCR was Performed with the Following Reaction Components:

[0600] 10 μl 10× Pfu buffer

[0601] 1 μl 10 mM dNTPs

[0602] 2 μl 10 μM of each primer

[0603] 83 μl of sterile water

[0604] 1.5 μl Pfu DNA polymerase (Stratagene, La Jolla, Calif.)

[0605] 50 ng DNA

[0606] PCR Amplification was Performed Using the Following Reaction Conditions:

[0607] 96° C. for 2 minutes, followed by 40 cycles of:

[0608] 96° C. for 20 seconds;

[0609] 68° C. for 15 seconds; and

[0610] 72° C. for 2 minutes and 30 seconds, followed by a final extension step of:

[0611] 72° C. for 4 minutes.

[0612] The PCR product was digested with EcoRI and cloned into pPDM His that had been cut with Eco 72I and EcoRI. The amino acid sequence for the UL50-His construct was confirmed, and is disclosed in SEQ ID NO:96. The construct was then transformed into BLR pLys and BLR Codon Plus RP cells.

[0613] Expression of HSV UL19 in E. coli:

[0614] The HSV UL19 coding region (the DNA sequence of which is disclosed in SEQ ID NO:84) was PCR amplified from clone pET17b with the following primers: (SEQ ID NO:106) PDM-453: 5′gccgctcctgcccgcgacccccc 3′ (SEQ ID NO:107) PDM-457: 5′ccagaattcattacagagacaggccctttagc 3′

[0615] The PCR was Performed with the Following Reaction Components:

[0616] 10 μl 10× Pfu buffer

[0617] 1 μl 10 mM dNTPs

[0618] 2 μl 10 μM of each primer

[0619] 83 μl of sterile water

[0620] 1.5 μl Pfu DNA polymerase (Stratagene, La Jolla, Calif.)

[0621] 50 ng DNA

[0622] PCR Amplification was Performed Using the Following Reaction Conditions:

[0623] 96° C. for 2 minutes, followed by 40 cycles of:

[0624] 96° C. for 20 seconds;

[0625] 70° C. for 15 seconds; and

[0626] 72° C. for 4 minutes, followed by a final extension step of:

[0627] 72° C. for 4 minutes.

[0628] The PCR product was digested with EcoRI and cloned into pPDM His that had been cut with Eco 72I and EcoRI. The amino acid sequence for the UL19-His construct was confirmed, and is disclosed in SEQ ID NO:95. The construct was then transformed into BLR pLys and BLR Codon Plus RP cells.

[0629] Expression of HSV UL47 in E. coli:

[0630] The HSV UL47 coding region (the DNA sequence of which is disclosed in SEQ ID NO:87) was PCR amplified using the following primers: (SEQ ID NO:108) PDM-631: 5′cactccgtggcgcgggcatgccg 3′ (SEQ ID NO:109) PDM-632: 5′ccgttagaattcactatgggcgtggcgggcc 3′

[0631] The PCR was Performed with the Following Reaction Components:

[0632] 10 μl 10× Pfu buffer

[0633] 1 μl 10 mM dNTPs

[0634] 2 μl 10 μM of each primer

[0635] 83 μl of sterile water

[0636] 1.5 μl Pfu DNA polymerase (Stratagene, La Jolla, Calif.)

[0637] 50 ng DNA

[0638] PCR Amplification was Performed Using the Following Reaction Conditions:

[0639] 96° C. for 2 minutes, followed by 40 cycles of:

[0640] 96° C. for 20 seconds;

[0641] 67° C. for 15 seconds; and

[0642] 72° C. for 2 minutes and 30 seconds, followed by a final extension step of:

[0643] 72° C. for 4 minutes.

[0644] The PCR product was digested with EcoRI and cloned into pPDM His that had been cut with Eco 72I and EcoRI. The amino acid sequence for the UL47-His construct was confirmed, and is disclosed in SEQ ID NO:94. The construct was then transformed into BLR pLys and BLR Codon Plus RP cells. Protein yields were low using this construct. UL47 was also cloned into PPDM Trx with two histadine tags that had been digested with StuI and EcoRI. The DNA and amino acid sequences for this construct are disclosed in SEQ ID NOs:86 and 92, respectively. Protein yields were much higher using this fusion construct.

[0645] Four additional fragments of UL47, designated UL47 A-D were also PCR amplified.

[0646] The UL47 A Coding Region was Amplified Using the Following Primer Pairs: (SEQ ID NO:110) PDM-631: 5′cactccgtgcgcgggcatgccg 3′ (SEQ ID NO:111) PDM-645: 5′catagaattcatcacgcgcgggaggggctggttttgc 3′

[0647] The UL47 B Coding Region was Amplified Using the Following Primer Pairs: (SEQ ID NO:112) PDM-646: 5′gacacggtggtcgcgtgcgtggc 3′ (SEQ ID NO:113) PDM-632: 5′ccgttagaattcactatgggcgtggcgggcc 3′.

[0648] Both Fragments were Amplified Using the Following PCR Reaction Components:

[0649] 10 μl 10× Pfu buffer

[0650] 1 μl 10 mM dNTPs

[0651] 2 μl 10 μM of each primer

[0652] 83 μl of sterile water

[0653] 1.5 μl Pfu DNA polymerase (Stratagene, La Jolla, Calif.)

[0654] 50 ng DNA

[0655] PCR Amplification was Performed Using the Following Reaction Conditions:

[0656] 96° C. for 2 minutes, followed by 40 cycles of:

[0657] 96° C. for 20 seconds;

[0658] 67° C. for 15 seconds; and

[0659] 72° C. for 2 minutes, followed by a final extension step of:

[0660] 72° C. for 4 minutes.

[0661] The UL47 C Coding Region was Amplified Using the Following Primer Pairs: (SEQ ID NO:114) PDM-631: 5′cactccgtgcgcgggcatgccg 3′ (SEQ ID NO:115) PDM-739: 5′cgtatgaattcatcagacccacccgttg 3′

[0662] The UL47 D Coding Region was Amplified Using the Following Primer Pairs: PDM-740: 5′gtgctggcgacggggctcatcc3′ (SEQ ID NO:116) PDM-632: 5′ccgttagaattcactatgggcgtg (SEQ ID NO:117) gcgggcc3′.

[0663] Both Fragments were Amplified Using the Following PCR Reaction Components:

[0664] 10 μl 10× Pfu buffer

[0665] 1 μl 10 mM dNTPs

[0666] 2 μl 10 μM of each primer

[0667] 83 μl of sterile water

[0668] 1.5 μl Pfu DNA polymerase (Stratagene, LaJolla, Calif.)

[0669] 50 ng DNA

[0670] PCR Amplification was Performed Using the Following Reaction Conditions:

[0671] 96° C. for 2 minutes, followed by 40 cycles of:

[0672] 96° C. for 20 seconds;

[0673] 63° C. for 15 seconds; and

[0674] 72° C. for 2 minutes, followed by a final extension step of:

[0675] 72° C. for 4 minutes.

[0676] The PCR product fromUL47 C was digested with EcoRI and cloned into pPDM His that had been digested with Eco 72I and EcoRI. The sequence was confirmed then the construct was transformed into BLR pLys S and BLR CodonPlus RP cells. The DNA and amino acid sequences of UL47 C are disclosed in SEQ ID NOs:88 and 93, respectively.

EXAMPLE 12 Identification of a Novel DNA Sequence Encoding the HSV-2 Gene, US8

[0677] The US8 gene of HSV-2 was cloned from the laboratory HG52 viral strain and sequenced, the DNA and amino acid sequences of which are disclosed in SEQ ID NOs:118 and 120, respectively. SEQ ID NO:118 was then compared to the HSV-2 HG52 strain genomic sequence contained in GenBank (accession number Z86099), the DNA and amino acid sequences of which are disclosed in SEQ ID NOs:119 and 121, respectively. This comparison revealed that SEQ ID NO:118 contained an extra base pair at position 542 that results in a frameshift. The presence of the extra base pair was also confirmed in a second laboratory strain of HSV-2, 333. There was one additional base pair (bp 156) upstream of the first stop codon in SEQ ID NO:118 that differed from the GenBank US8 sequence (SEQ ID NO:119). No change in the US8 amino acid sequence would result from the change in the nucleotide sequence at base pair 156.

[0678] In addition to examining the sequence of a number of laboratory strains of HSV-2, genomic DNA sequence was also obtained from two clinically isolated viral samples, donors RW1874 and HV5101). Using PCR primers designed to gene specific sequences both up- and down-stream of the position 542 insertion, this region was PCR amplified and directly sequenced from the purified amplicon using the same primer pair. The sequences obtained from both RW1874 and HV5101 showed the additional guanine nucleotide at position 542. HV5101 had one additional base pair change at base pair 571 (G/571/C:HV5101/location/HG52) when compared to HG52 (SEQ ID NO:119). This difference is a non-conservative change in the frameshift form.

EXAMPLE 13 Vaccination with the HSV-2 UL47 Protein Elicits both a CD4⁺ and CD8⁺ Specific T Cell Response

[0679] This example demonstrates the effectiveness of UL47 as a vaccine against HSV-2. Balb/c mice vaccinated with UL47, delivered by plasmid DNA, mounted a UL47-specific CD4⁺ and CD8⁺ cell response.

[0680] Two Balb/c mice were immunized three times with 100 μg of UL47 plasmid DNA (UL47 DNA), an additional four mice were immunized twice with UL47, followed by infection with 1×10³ pfu of an attenuated HSV-2 strain, 333vhsB (UL47 DNA/HSV). A further four mice received HSV-2 infection alone (HSV control). The spleens were harvested two weeks post-final immunization and stimulated in vitro with vaccinia-UL47 for 7 days.

[0681] On day 7, the splenocytes were assayed for cytotoxic activity by chromium release against P815 cells pulsed with pools of 10-15-mer peptides that spanned the UL47 gene (18 pools total). The splenocytes were re-stimulated in vitro and then re-assayed against positive peptide pools, plus the constitutive 15-mer peptides. At an effector:target ratio of 100:1, specific lytic activity by CD8+cells could be seen in response to P815 cells pulsed with peptides 85 (SEQ ID NO:122), 89 (SEQ ID NO:123), 99 and 98 (SEQ ID NO:124), 105 (SEQ ID NO:125), and 112 (SEQ ID NO:126).

[0682] In order to determine the presence of a CD4+T cell responses, splenocytes were stimulated in vitro with 5 μg/ml recombinant UL47 (rUL47). Three days following stimulation, the culture supernatants were harvested and assayed for IFN-gamma by ELISA. Supernatants harvested from both the splenocytes from the “UL47 DNA” mice (those that were immunized) and the “UL47 DNA/HSV” mice (those that were immunized followed by infection with HSV) had significant levels of IFN-gamma present compared to the “HSV control” mice (those who were uninmmunized and infected).

[0683] A further four mice were immunized four times with UL47 DNA and their splenocytes harvested. The splenocytes were then stimulated with peptides p85, p89, p98, p99, p105, and p112 and the CD8+cells assayed for the presence of intracellular IFN-gamma production using flow cytometry. The percentages of CD8+ cells producing IFN-gamma were significant in the splenocytes stimulated with peptides p85, p89, p98, p99, p105 and p112, compared to the control cells (cells stimulated with media or PBS alone). Reponses seen against peptides p98 and p99 should the highest percentages, with greater than 2% of all CD8+ splenocytes positive for intracellular IFN-gamma.

[0684] These data further demonstrate the effectiveness of UL47 as a vaccine candidate in the protection against or treatment of HSV infection.

EXAMPLE 14 CD8+ T Cell Responses from HSV-2 Seropositive Donors

[0685] Six HSV-2 seropositive donors were screened to determine which HSV-2 proteins were capable of eliciting a CD8+T cell response. The donors included: AD104, AD116, AD120, D477, HV5101, and JH6376. In order to determine which HSV-2 proteins were immunogenic, synthetic peptides (15-mers overlapping by 11 amino acids) were synthesized across the following region of several HSV-2 polypeptides, including: UL15 (a.a. 600-734), UL18 (a.a.1-110), UL23 (a.a. 241-376), UL46 (a.a. 617-722), UL47 (a.a. 1-696), UL49 (a.a. 1-300), ICP27 (a.a. 1-512), US3 (a.a. 125-276), and US8A (a.a. 83-146). Peptides synthesized for UL47, UL49, and ICP27 spanned the full-length polypeptide. Peptides synthesized for UL15, UL18, UL23, UL46, US3, and US8A spanned the portions of these polypeptides previously determined to encode antigens recognized by CD4⁺ T cells during CD4 expression-cloning library screening.

[0686] The donors CD8⁺ T cells were isolated from PBMC using the following procedure: initially peripheral blood lymphocytes (PBL) were separate from macrophages using plastic adherence. The CD8⁺ T cells were then further purified by depletion of non-CD8⁺ cells using a commercial MACS bead kit (Miltenyi). CD8⁺ T cells isolated using this method are generally >95% CD8⁺/CD3⁺/CD4⁻, as measured by flow cytometry (FACS). Peptides were screened by 24-hour co-culture of CD8⁺ T cells (2×10⁵/well), autologous dendritic cells (1×10⁴/well), and peptides (10 μg/ml each) in 96 well ELISPOT plates pre-coated with anti-human IFN-gamma antibody. Peptides were initially screened as pools of ≧10 peptides. ELISPOT plates were subsequently developed per a standard protocol. The numbers of spots per well were counted using an automated video-microscopy ELISPOT reader. Peptide from pools screening positive were subsequently tested individually in a second ELISPOT assay.

[0687] For AD104, only the peptide US3 #33 (SEQ ID NO:139: amino acids 262-276) scored positive.

[0688] For AD116, peptides UL15 #23 (SEQ ID NO:127: amino acids 688-702), UL15 #30 (SEQ ID NO:128: amino acids 716-730), UL23 #7 (SEQ ID NO:129: amino acids 265-272), UL46 #2 (SEQ ID NO:130: amino acids 621-635), UL46 #8 (SEQ ID NO:131: amino acids 645-659), UL46 #9 (SEQ ID NO:132: amino acids 649-663), UL46 #11 (SEQ ID NO:133: amino acids 657-671), UL47 #86 (SEQ ID NO:134: amino acids 341-355), UL49 #6 (SEQ ID NO:135: amino acids 21-35), UL49 #49 (SEQ ID NO:138: amino acids 193-208), and US8A #5 (SEQ ID NO:140: amino acids 99-113) scored positive both pooled and individually. In addition, AD116 also recognized the B*0702-restricted epitope UL49 #12 (SEQ ID NO:136: amino acids 45-59) and UL49 #13 (SEQ ID NO:137: amino acids 49 to 63).

[0689] Donors D477, HV5101, and JH6376 T cells recognized the HLA-A*0201-restricted epitopes UL47 #73/#74 (amino acids 289-297) and UL47 #137/#138 (amino acids 550-559), respectively.

[0690] Donor AD120 scored positive for one peptide pool, UL46 #1-12.

[0691] Donor D477 scored positive for 5 peptide pools: UL18 #1-12, UL23 #1-10, UL23 #11-20, UL46 #1-12, and UL49 #11-20.

EXAMPLE 15 Identification of a Novel Sequence Coding for the US4 Protein of HSV-2

[0692] Screening the HSV2-11 library with T cells from donor AD104 had previously identified the clone insert F10B3 (see Example 1 for details). SEQ ID NO:8, corresponds to the partial sequence of the insert from clone HSV2II_US3/US4 fragF10B3_T7Trc.seq, and contains a potential open reading frame having an amino acid sequence set forth in SEQ ID NO: 10. The full-length DNA and amino acid sequences corresponding to the insert sequence are disclosed in SEQ ID NOs:141 and 142, respectively. The full length US4 HG52 DNA and amino acid sequence are disclosed in SEQ ID NO:179 and 143, respectively, and differs from the insert sequence as follows: S35N (HG52/location/333).

EXAMPLE 16 Identification of HSV-2-Specific CD8+ T Cell Responses in HSV-2

[0693] CD8⁺ T cells isolated from a panel of HSV-2 seropositive donors were screened for their ability to respond to a variety of HSV-2 proteins. Briefly, PBMCs were obtained from donors EB5491, AG10295, LM10295, and 447, and enriched for CD8⁺ T cells using microbeads or CD8+ Enrichment Kits from Miltenyi. Synthetic peptides (15 amino acids in length and overlapping in sequence by 10 or 11 amino acids) were synthesized across several complete or partial ORFs from HSV-2 strain HG52, including proteins UL21 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.144 and 154, respectively), UL50 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.145 and 153, respectively), US3 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.146 and 154, respectively), UL54 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.147 and 156, respectively), US8 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.148 and 157, respectively), UL19 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.149 and 158, respectively), UL46 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.150 and 159, respectively), UL18 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.151 and 160, respectively), and RL2 (the full length DNA/amino acids of which are disclosed in SEQ ID NOs:.152 and 161, respectively). The peptides were screened by 24 co-culture of the donor's CD8+T cells (2-5×10⁵ cells/well), autologous dendritic cells (2-5×10⁴ cells/well) and peptides (0.5 μg/ml each) in 96-well ELISPOT plates that had been pre-coated with anti-human IFN-Y antibody. Each peptide pool was screened in an individual well. The ELISPOT plates were developed as per a standard protocol. The number of spots per well was counted using an automated video-microscopy ELISPOT reader. Individual 15-mer peptides, determined from peptide pools testing positive, were screened as described above and returned the following results:

[0694] Donor EB5491 demonstrated CD8+T cell responses to the HSV-2 antigens: ICP0 peptide #43 (amino acids 211-225: IWTGNPRTAPRSLSL: SEQ ID NO:162). UL46 peptides #41 (amino acids 201-215: YMFFMRPADPSRPST: SEQ ID NO:163), UL46 #50 (amino acids 246-260: VCRRLGPADRRFVAL: SEQ ID NO:164), UL46 #51 (amino acids 251-265: GPADRRFVALSGSLE: SEQ ID NO:165), and UL46 #60 (amino acids 296-310: SDVLGHLTRLAHLWE: SEQ ID NO:166). Donor EB5491 also demonstrated a CD8+T cell response to the HSV-2 protein, US8 #74 (amino acids 366-380: HGMTISTMQYRNAV: SEQ ID NO:167).

[0695] Donor JH6376 demonstrated CD8+ T cells responses to the HSV-2 proteins ICP0, which corresponded to a 9-mer mapped to amino acids 215-223 (NPRTAPRSL: SEQ ID NO:177) and UL46, which corresponded to a 10-mer mapped to amino acids 251-260 (GPADRRFVAL: SEQ ID NO:178).

[0696] Donor AG1059 demonstrated CD8+ T cell responses to the HSV-2 proteins UL19 peptide 102 (amino acids 506-520: LNAWRQRLAHGRVRW: SEQ ID NO:168), UL19 #103 (amino acids 511-525: QRLAHGRVRWVAECQ: SEQ ID NO:169) and UL18 #17 (amino acids 65-79: LAYRRRFPAVITRVL: SEQ ID NO:172) and UL18 #18 (amino acids 69-83: RRFPAVITRVLPTRI: SEQ ID NO:173).

[0697] Donor LM10295 demonstrated CD8+ T cell responses to the HSV-2 protein UL19 #74 (amino acids 366-380: DLVAIGDRLVFLEAL: SEQ ID NO:170) and UL19 #75 (amino acids 371-385: GDRLVFLEALERRIY: SEQ ID NO:171).

[0698] Donor 477 demonstrated CD8+ T cell responses to the HSV-2 protein UL50 #16 (amino acids 76-90: CAIIHAPAVSGPGPH: SEQ ID NO:174), UL50 #23 (amino acids 111-125: PNGTRGFAPGALRVD: SEQ ID NO:175), and UL50 #49 (amino acids 241-255: LRVLRAADGPEACYV: SEQ ID NO:176).

EXAMPLE 17 Expression of a Truncated form of UL47 in E. coli

[0699] A C-terminal truncation of the full length UL47 coding region was expressed in E. coli, and designated as UL47F. This truncated portion of UL47 contains the C-terminal T cell epitope of UL47, corresponding to amino acids 500-559.

[0700] Expression of HSV UL47 F in E. coli:

[0701] The HSV UL47F coding region (the DNA and amino acid sequences of which are disclosed in SEQ ID NO:180 and 181, respectively) was PCR amplified using the following primers: CBH-631: 5′ctgggtctggctgacacggtggtc (SEQ ID NO:182) gcgtgcgtg3′ PDM-632: 5′ccgttagaattcactatgggcgtg (SEQ ID NO:183) gcgggcc3′

[0702] The PCR was Performed With the Following Reaction Components:

[0703] 10 μl 10× Pfu buffer

[0704] 1 μl 10 mM dNTPs

[0705] 2 μl 10 μM of each primer

[0706] 83 μl of sterile water

[0707] 1.5 μl Pfu DNA polymerase (Stratagene, La Jolla, Calif.)

[0708] 50 ng DNA

[0709] PCR Amplification was Performed Using the Following Reaction Conditions:

[0710] 96° C. for 2 minutes, followed by 40 cycles of:

[0711] 96° C. for 20 seconds;

[0712] 68° C. for 15 seconds; and

[0713] 72° C. for 1 minute and 30 seconds, followed by a final extension step of:

[0714] 72° C. for 4 minutes.

[0715] The PCR product was digested with EcoRI and cloned into pPDM His that had been cut with Eco 72I and EcoRI. The sequence of the construct was confirmed, and then the construct was transformed into BRL pLys S and BLR CodonPlus RP cells.

[0716] Those skilled in the art will appreciate that the conceptions and specific embodiments disclosed in the foregoing description may be readily utilized as a basis for modifying or designing other embodiments for carrying out the same purposes of the present invention. Those skilled in the art will also appreciate that such equivalent embodiments do not depart from the spirit and scope of the invention as set forth in the appended claims.

[0717] From the foregoing it will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims.

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 183 <210> SEQ ID NO 1 <211> LENGTH: 815 <212> TYPE: DNA <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 1 ccacgccgcc gcaccccagg cggacgtggc gccggttctg gacagccagc ccactgtggg 60 aacggacccc ggctacccag tccccctaga actcacgccc gagaacgcgg aggcggtggc 120 gcggtttctg ggggacgccg tcgaccgcga gcccgcgctc atgctggagt acttctgtcg 180 gtgcgcccgc gaggagagca agcgcgtgcc cccacgaacc ttcggcagcg ccccccgcct 240 cacggaggac gactttgggc tcctgaacta cgcgctcgct gagatgcgac gcctgtgcct 300 ggaccttccc ccggtccccc ccaacgcata cacgccctat catctgaggg agtatgcgac 360 gcggctggtt aacgggttca aacccctggt gcggcggtcc gcccgcctgt atcgcatcct 420 ggggattctg gttcacctgc gcatccgtac ccgggaggcc tcctttgagg aatggatgcg 480 ctccaaggag gtggacctgg acttcgggct gacggaaagg cttcgcgaac acgaggccca 540 gctaatgatc ctggcccagg ccctgaaccc ctacgactgt ctgatccaca gcaccccgaa 600 cacgctcgtc gagcgggggc tgcagtcggc gctgaagtac gaagagtttt acctcaagcg 660 cttcggcggg cactacatgg agtccgtctt ccagatgtac acccgcatcg ccgggttcct 720 ggcgtgccgg gcgacccgcg gcatgcgcca catcgccctg gggcgacagg ggtcgtggtg 780 ggaaatgttc aagttctttt tccaccgcct ctacg 815 <210> SEQ ID NO 2 <211> LENGTH: 271 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 2 His Ala Ala Ala Pro Gln Ala Asp Val Ala Pro Val Leu Asp Ser Gln 1 5 10 15 Pro Thr Val Gly Thr Asp Pro Gly Tyr Pro Val Pro Leu Glu Leu Thr 20 25 30 Pro Glu Asn Ala Glu Ala Val Ala Arg Phe Leu Gly Asp Ala Val Asp 35 40 45 Arg Glu Pro Ala Leu Met Leu Glu Tyr Phe Cys Arg Cys Ala Arg Glu 50 55 60 Glu Ser Lys Arg Val Pro Pro Arg Thr Phe Gly Ser Ala Pro Arg Leu 65 70 75 80 Thr Glu Asp Asp Phe Gly Leu Leu Asn Tyr Ala Leu Ala Glu Met Arg 85 90 95 Arg Leu Cys Leu Asp Leu Pro Pro Val Pro Pro Asn Ala Tyr Thr Pro 100 105 110 Tyr His Leu Arg Glu Tyr Ala Thr Arg Leu Val Asn Gly Phe Lys Pro 115 120 125 Leu Val Arg Arg Ser Ala Arg Leu Tyr Arg Ile Leu Gly Ile Leu Val 130 135 140 His Leu Arg Ile Arg Thr Arg Glu Ala Ser Phe Glu Glu Trp Met Arg 145 150 155 160 Ser Lys Glu Val Asp Leu Asp Phe Gly Leu Thr Glu Arg Leu Arg Glu 165 170 175 His Glu Ala Gln Leu Met Ile Leu Ala Gln Ala Leu Asn Pro Tyr Asp 180 185 190 Cys Leu Ile His Ser Thr Pro Asn Thr Leu Val Glu Arg Gly Leu Gln 195 200 205 Ser Ala Leu Lys Tyr Glu Glu Phe Tyr Leu Lys Arg Phe Gly Gly His 210 215 220 Tyr Met Glu Ser Val Phe Gln Met Tyr Thr Arg Ile Ala Gly Phe Leu 225 230 235 240 Ala Cys Arg Ala Thr Arg Gly Met Arg His Ile Ala Leu Gly Arg Gln 245 250 255 Gly Ser Trp Trp Glu Met Phe Lys Phe Phe Phe His Arg Leu Tyr 260 265 270 <210> SEQ ID NO 3 <211> LENGTH: 1142 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 3 Met Ala Asn Arg Pro Ala Ala Ser Ala Leu Ala Gly Ala Arg Ser Pro 1 5 10 15 Ser Glu Arg Gln Glu Pro Arg Glu Pro Glu Val Ala Pro Pro Gly Gly 20 25 30 Asp His Val Phe Cys Arg Lys Val Ser Gly Val Met Val Leu Ser Ser 35 40 45 Asp Pro Pro Gly Pro Ala Ala Tyr Arg Ile Ser Asp Ser Ser Phe Val 50 55 60 Gln Cys Gly Ser Asn Cys Ser Met Ile Ile Asp Gly Asp Val Ala Arg 65 70 75 80 Gly His Leu Arg Asp Leu Glu Gly Ala Thr Ser Thr Gly Ala Phe Val 85 90 95 Ala Ile Ser Asn Val Ala Ala Gly Gly Asp Gly Arg Thr Ala Val Val 100 105 110 Ala Leu Gly Gly Thr Ser Gly Pro Ser Ala Thr Thr Ser Val Gly Thr 115 120 125 Gln Thr Ser Gly Glu Phe Leu His Gly Asn Pro Arg Thr Pro Glu Pro 130 135 140 Gln Gly Pro Gln Ala Val Pro Pro Pro Pro Pro Pro Pro Phe Pro Trp 145 150 155 160 Gly His Glu Cys Cys Ala Arg Arg Asp Ala Arg Gly Gly Ala Glu Lys 165 170 175 Asp Val Gly Ala Ala Glu Ser Trp Ser Asp Gly Pro Ser Ser Asp Ser 180 185 190 Glu Thr Glu Asp Ser Asp Ser Ser Asp Glu Asp Thr Gly Ser Glu Thr 195 200 205 Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala Gly Ala Thr Asp Asp Asp 210 215 220 Asp Ser Asp Ser Asp Ser Arg Ser Asp Asp Ser Val Gln Pro Asp Val 225 230 235 240 Val Val Arg Arg Arg Trp Ser Asp Gly Pro Ala Pro Val Ala Phe Pro 245 250 255 Lys Pro Arg Arg Pro Gly Asp Ser Pro Gly Asn Pro Gly Leu Gly Ala 260 265 270 Gly Thr Gly Pro Gly Ser Ala Thr Asp Pro Arg Ala Ser Ala Asp Ser 275 280 285 Asp Ser Ala Ala His Ala Ala Ala Pro Gln Ala Asp Val Ala Pro Val 290 295 300 Leu Asp Ser Gln Pro Thr Val Gly Thr Asp Pro Gly Tyr Pro Val Pro 305 310 315 320 Leu Glu Leu Thr Pro Glu Asn Ala Glu Ala Val Ala Arg Phe Leu Gly 325 330 335 Asp Ala Val Asp Arg Glu Pro Ala Leu Met Leu Glu Tyr Phe Cys Arg 340 345 350 Cys Ala Arg Glu Glu Ser Lys Arg Val Pro Pro Arg Thr Phe Gly Ser 355 360 365 Ala Pro Arg Leu Thr Glu Asp Asp Phe Gly Leu Leu Asn Tyr Ala Leu 370 375 380 Ala Glu Met Arg Arg Leu Cys Leu Asp Leu Pro Pro Val Pro Pro Asn 385 390 395 400 Ala Tyr Thr Pro Tyr His Leu Arg Glu Tyr Ala Thr Arg Leu Val Asn 405 410 415 Gly Phe Lys Pro Leu Val Arg Arg Ser Ala Arg Leu Tyr Arg Ile Leu 420 425 430 Gly Val Leu Val His Leu Arg Ile Arg Thr Arg Glu Ala Ser Phe Glu 435 440 445 Glu Trp Met Arg Ser Lys Glu Val Asp Leu Asp Phe Gly Leu Thr Glu 450 455 460 Arg Leu Arg Glu His Glu Ala Gln Leu Met Ile Leu Ala Gln Ala Leu 465 470 475 480 Asn Pro Tyr Asp Cys Leu Ile His Ser Thr Pro Asn Thr Leu Val Glu 485 490 495 Arg Gly Leu Gln Ser Ala Leu Lys Tyr Glu Glu Phe Tyr Leu Lys Arg 500 505 510 Phe Gly Gly His Tyr Met Glu Ser Val Phe Gln Met Tyr Thr Arg Ile 515 520 525 Ala Gly Phe Leu Ala Cys Arg Ala Thr Arg Gly Met Arg His Ile Ala 530 535 540 Leu Gly Arg Gln Gly Ser Trp Trp Glu Met Phe Lys Phe Phe Phe His 545 550 555 560 Arg Leu Tyr Asp His Gln Ile Val Pro Ser Thr Pro Ala Met Leu Asn 565 570 575 Leu Gly Thr Arg Asn Tyr Tyr Thr Ser Ser Cys Tyr Leu Val Asn Pro 580 585 590 Gln Ala Thr Thr Asn Gln Ala Thr Leu Arg Ala Ile Thr Gly Asn Val 595 600 605 Ser Ala Ile Leu Ala Arg Asn Gly Gly Ile Gly Leu Cys Met Gln Ala 610 615 620 Phe Asn Asp Ala Ser Pro Gly Thr Ala Ser Ile Met Pro Ala Leu Lys 625 630 635 640 Val Leu Asp Ser Leu Val Ala Ala His Asn Lys Gln Ser Thr Arg Pro 645 650 655 Thr Gly Ala Cys Val Tyr Leu Glu Pro Trp His Ser Asp Val Arg Ala 660 665 670 Val Leu Arg Met Lys Gly Val Leu Ala Gly Glu Glu Ala Gln Arg Cys 675 680 685 Asp Asn Ile Phe Ser Ala Leu Trp Met Pro Asp Leu Phe Phe Lys Arg 690 695 700 Leu Ile Arg His Leu Asp Gly Glu Lys Asn Val Thr Trp Ser Leu Phe 705 710 715 720 Asp Arg Asp Thr Ser Met Ser Leu Ala Asp Phe His Gly Glu Glu Phe 725 730 735 Glu Lys Leu Tyr Glu His Leu Glu Ala Met Gly Phe Gly Glu Thr Ile 740 745 750 Pro Ile Gln Asp Leu Ala Tyr Ala Ile Val Arg Ser Ala Ala Thr Thr 755 760 765 Gly Ser Pro Phe Ile Met Phe Lys Asp Ala Val Asn Arg His Tyr Ile 770 775 780 Tyr Asp Thr Gln Gly Ala Ala Ile Ala Gly Ser Asn Leu Cys Thr Glu 785 790 795 800 Ile Val His Pro Ala Ser Lys Arg Ser Ser Gly Val Cys Asn Leu Gly 805 810 815 Ser Val Asn Leu Ala Arg Cys Val Ser Arg Gln Thr Phe Asp Phe Gly 820 825 830 Arg Leu Arg Asp Ala Val Gln Ala Cys Val Leu Met Val Asn Ile Met 835 840 845 Ile Asp Ser Thr Leu Gln Pro Thr Pro Gln Cys Thr Arg Gly Asn Asp 850 855 860 Asn Leu Arg Ser Met Gly Ile Gly Met Gln Gly Leu His Thr Ala Cys 865 870 875 880 Leu Lys Met Gly Leu Asp Leu Glu Ser Ala Glu Phe Arg Asp Leu Asn 885 890 895 Thr His Ile Ala Glu Val Met Leu Leu Ala Ala Met Lys Thr Ser Asn 900 905 910 Ala Leu Cys Val Arg Gly Ala Arg Pro Phe Ser His Phe Lys Arg Ser 915 920 925 Met Tyr Arg Ala Gly Arg Phe His Trp Glu Arg Phe Ser Asn Ala Ser 930 935 940 Pro Arg Tyr Glu Gly Glu Trp Glu Met Leu Arg Gln Ser Met Met Lys 945 950 955 960 His Gly Leu Arg Asn Ser Gln Phe Ile Ala Leu Met Pro Thr Ala Ala 965 970 975 Ser Ala Gln Ile Ser Asp Val Ser Glu Gly Phe Ala Pro Leu Phe Thr 980 985 990 Asn Leu Phe Ser Lys Val Thr Arg Asp Gly Glu Thr Leu Arg Pro Asn 995 1000 1005 Thr Leu Leu Leu Lys Glu Leu Glu Arg Thr Phe Gly Gly Lys Arg Leu 1010 1015 1020 Leu Asp Ala Met Asp Gly Leu Glu Ala Lys Gln Trp Ser Val Ala Gln 1025 1030 1035 1040 Ala Leu Pro Cys Leu Asp Pro Ala His Pro Leu Arg Arg Phe Lys Thr 1045 1050 1055 Ala Phe Asp Tyr Asp Gln Glu Leu Leu Ile Asp Leu Cys Ala Asp Arg 1060 1065 1070 Ala Pro Tyr Val Asp His Ser Gln Ser Met Thr Leu Tyr Val Thr Glu 1075 1080 1085 Lys Ala Asp Gly Thr Leu Pro Ala Ser Thr Leu Val Arg Leu Leu Val 1090 1095 1100 His Ala Tyr Lys Arg Gly Leu Lys Thr Gly Met Tyr Tyr Cys Lys Val 1105 1110 1115 1120 Arg Lys Ala Thr Asn Ser Gly Val Phe Ala Gly Asp Asp Asn Ile Val 1125 1130 1135 Cys Thr Ser Cys Ala Leu 1140 <210> SEQ ID NO 4 <211> LENGTH: 208 <212> TYPE: DNA <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 4 gcgccgcgcc cgcgtgccgc agaccacctc gcggcggctc ccccgcggcc tttcccgtgg 60 ccctccacgc cgtggacgcc ccctcccaat tcgtcacctg gctcgccgtg cgctggctgc 120 ggggggcggt gggtctcggg gccgtcctgt gcgggattgc gttttacgtg acgtcaatcg 180 cccgaggcgc ataaaggtcc ggcggcca 208 <210> SEQ ID NO 5 <211> LENGTH: 64 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 5 Gly Ala Ala Pro Ala Cys Arg Arg Pro Pro Arg Gly Gly Ser Pro Ala 1 5 10 15 Ala Phe Pro Val Ala Leu His Ala Val Asp Ala Pro Ser Gln Phe Val 20 25 30 Thr Trp Leu Ala Val Arg Trp Leu Arg Gly Ala Val Gly Leu Gly Ala 35 40 45 Val Leu Cys Gly Ile Ala Phe Tyr Val Thr Ser Ile Ala Arg Gly Ala 50 55 60 <210> SEQ ID NO 6 <211> LENGTH: 70 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 6 Arg Arg Ala Arg Val Pro Gln Thr Thr Ser Arg Arg Leu Pro Arg Gly 1 5 10 15 Leu Ser Arg Gly Pro Pro Arg Arg Gly Arg Pro Leu Pro Ile Arg His 20 25 30 Leu Ala Arg Arg Ala Leu Ala Ala Gly Gly Gly Gly Ser Arg Gly Arg 35 40 45 Pro Val Arg Asp Cys Val Leu Arg Asp Val Asn Arg Pro Arg Arg Ile 50 55 60 Lys Val Arg Arg Pro Ala 65 70 <210> SEQ ID NO 7 <211> LENGTH: 146 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 7 Met Asp Pro Ala Leu Arg Ser Tyr His Gln Arg Leu Arg Leu Tyr Thr 1 5 10 15 Pro Ile Ala Arg Gly Val Asn Leu Ala Ala Arg Ser Pro Pro Leu Val 20 25 30 Arg Glu Ala Arg Ala Val Val Thr Pro Arg Pro Pro Ile Arg Pro Ser 35 40 45 Ser Gly Lys Ala Ser Ser Asp Asp Ala Asp Val Gly Asp Glu Leu Ile 50 55 60 Ala Ile Ala Asp Ala Arg Gly Asp Pro Pro Glu Thr Leu Pro Pro Gly 65 70 75 80 Ala Gly Gly Ala Ala Pro Ala Cys Arg Arg Pro Pro Arg Gly Gly Ser 85 90 95 Pro Ala Ala Phe Pro Val Ala Leu His Ala Val Asp Ala Pro Ser Gln 100 105 110 Phe Val Thr Trp Leu Ala Val Arg Trp Leu Arg Gly Ala Val Gly Leu 115 120 125 Gly Ala Val Leu Cys Gly Ile Ala Phe Tyr Val Thr Ser Ile Ala Arg 130 135 140 Gly Ala 145 <210> SEQ ID NO 8 <211> LENGTH: 137 <212> TYPE: DNA <213> ORGANISM: Herpes simplx virus <400> SEQUENCE: 8 ccccaccgcc cccccacagg cggcgcgtgc ggagggcggc ccgtgcgtcc ccccggtccc 60 cgcgggccgc ccgtggcgct cggtgccccc ggtatggtat tccgccccca accccgggtt 120 tcgtggcctg cgtttcc 137 <210> SEQ ID NO 9 <211> LENGTH: 430 <212> TYPE: DNA <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 9 atggaccggg aggcacttcg ggccatcagc cgcgggtgca agcccccttc gaccctggca 60 aaactggtga ccgggctggg attcgcgatc cacggagcgc tcatcccggg gtcggagggg 120 tgtgtctttg atagcagcca cccgaactac cctcatcggg taatcgtcaa ggcggggtgg 180 tacgccagca cgaaccacga ggcgcggctg ctgagacgcc tgaaccaccc cgcgatccta 240 cccctcctgg acctgcacgt cgtttctggg gtcacgtgtc tggtcctccc caagtatcac 300 tgcgacctgt atacctatct gagcaagcgc ccgtctccgt tgggccacct acagataacc 360 gcggtctccc ggcagctctt gagcgccatc gactacgtcc actgcgaagg catcatccac 420 cgcgatatta 430 <210> SEQ ID NO 10 <211> LENGTH: 22 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 10 Trp Thr Gly Arg His Phe Gly Pro Ser Ala Ala Gly Ala Ser Pro Leu 1 5 10 15 Arg Pro Trp Gln Asn Trp 20 <210> SEQ ID NO 11 <211> LENGTH: 143 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 11 Met Asp Arg Glu Ala Leu Arg Ala Ile Ser Arg Gly Cys Lys Pro Pro 1 5 10 15 Ser Thr Leu Ala Lys Leu Val Thr Gly Leu Gly Phe Ala Ile His Gly 20 25 30 Ala Leu Ile Pro Gly Ser Glu Gly Cys Val Phe Asp Ser Ser His Pro 35 40 45 Asn Tyr Pro His Arg Val Ile Val Lys Ala Gly Trp Tyr Ala Ser Thr 50 55 60 Asn His Glu Ala Arg Leu Leu Arg Arg Leu Asn His Pro Ala Ile Leu 65 70 75 80 Pro Leu Leu Asp Leu His Val Val Ser Gly Val Thr Cys Leu Val Leu 85 90 95 Pro Lys Tyr His Cys Asp Leu Tyr Thr Tyr Leu Ser Lys Arg Pro Ser 100 105 110 Pro Leu Gly His Leu Gln Ile Thr Ala Val Ser Arg Gln Leu Leu Ser 115 120 125 Ala Ile Asp Tyr Val His Cys Glu Gly Ile Ile His Arg Asp Ile 130 135 140 <210> SEQ ID NO 12 <211> LENGTH: 481 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 12 Met Ala Cys Arg Lys Phe Cys Gly Val Tyr Arg Arg Pro Asp Lys Arg 1 5 10 15 Gln Glu Ala Ser Val Pro Pro Glu Thr Asn Thr Ala Pro Ala Phe Pro 20 25 30 Ala Ser Thr Phe Tyr Thr Pro Ala Glu Asp Ala Tyr Leu Ala Pro Gly 35 40 45 Pro Pro Glu Thr Ile His Pro Ser Arg Pro Pro Ser Pro Gly Glu Ala 50 55 60 Ala Arg Leu Cys Gln Leu Gln Glu Ile Leu Ala Gln Met His Ser Asp 65 70 75 80 Glu Asp Tyr Pro Ile Val Asp Ala Ala Gly Ala Glu Glu Glu Asp Glu 85 90 95 Ala Asp Asp Asp Ala Pro Asp Asp Val Ala Tyr Pro Glu Asp Tyr Ala 100 105 110 Glu Gly Arg Phe Leu Ser Met Val Ser Ala Ala Pro Leu Pro Gly Ala 115 120 125 Ser Gly His Pro Pro Val Pro Gly Arg Ala Ala Pro Pro Asp Val Arg 130 135 140 Thr Cys Asp Thr Gly Lys Val Gly Ala Thr Gly Phe Thr Pro Glu Glu 145 150 155 160 Leu Asp Thr Met Asp Arg Glu Ala Leu Arg Ala Ile Ser Arg Gly Cys 165 170 175 Lys Pro Pro Ser Thr Leu Ala Lys Leu Val Thr Gly Leu Gly Phe Ala 180 185 190 Ile His Gly Ala Leu Ile Pro Gly Ser Glu Gly Cys Val Phe Asp Ser 195 200 205 Ser His Pro Asn Tyr Pro His Arg Val Ile Val Lys Ala Gly Trp Tyr 210 215 220 Ala Ser Thr Ser His Glu Ala Arg Leu Leu Arg Arg Leu Asn His Pro 225 230 235 240 Ala Ile Leu Pro Leu Leu Asp Leu His Val Val Ser Gly Val Thr Cys 245 250 255 Leu Val Leu Pro Lys Tyr His Cys Asp Leu Tyr Thr Tyr Leu Ser Lys 260 265 270 Arg Pro Ser Pro Leu Gly His Leu Gln Ile Thr Ala Val Ser Arg Gln 275 280 285 Leu Leu Ser Ala Ile Asp Tyr Val His Cys Lys Gly Ile Ile His Arg 290 295 300 Asp Ile Lys Thr Glu Asn Ile Phe Ile Asn Thr Pro Glu Asn Ile Cys 305 310 315 320 Leu Gly Asp Phe Gly Ala Ala Cys Phe Val Arg Gly Cys Arg Ser Ser 325 330 335 Pro Phe His Tyr Gly Ile Ala Gly Thr Ile Asp Thr Asn Ala Pro Glu 340 345 350 Val Leu Ala Gly Asp Pro Tyr Thr Gln Val Ile Asp Ile Trp Ser Ala 355 360 365 Gly Leu Val Ile Phe Glu Thr Ala Val His Thr Ala Ser Leu Phe Ser 370 375 380 Ala Pro Arg Asp Pro Glu Arg Arg Pro Cys Asp Asn Gln Ile Ala Arg 385 390 395 400 Ile Ile Arg Gln Ala Gln Val His Val Asp Glu Phe Pro Thr His Ala 405 410 415 Glu Ser Arg Leu Thr Ala His Tyr Arg Ser Arg Ala Ala Gly Asn Asn 420 425 430 Arg Pro Ala Trp Thr Arg Pro Ala Trp Thr Arg Tyr Tyr Lys Ile His 435 440 445 Thr Asp Val Glu Tyr Leu Ile Cys Lys Ala Leu Thr Phe Asp Ala Ala 450 455 460 Leu Arg Pro Ser Ala Ala Glu Leu Leu Arg Leu Pro Leu Phe His Pro 465 470 475 480 Lys <210> SEQ ID NO 13 <211> LENGTH: 501 <212> TYPE: DNA <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 13 gggggcgcgt ctacgaggag atcccctggg ttcgggtata cgaaaacatc tgccttcgcc 60 ggcaagacgc cggcggggcg gccccgccgg gagacgcccc ggactccccg tacatcgagg 120 cggaaaatcc cctgtacgac tggggcgggt ctgccctctt ctcccctccg ggggccacac 180 gcgccccgga cccgggacta agcctgtcgc ccatgcccgc ccgcccccgg accaacgcgc 240 tggccaacga cggcccgaca aacgtcgccg ccctcagcgc cctgttgacg aagctcaaac 300 gcggccgaca ccagagccat taaaaaaatg cgaccgccgg ccccaccgtc tcggtttccg 360 gcccctttcc ccgtatgtct gttttcaata aaaagtaaca aacagagaaa aaaaaacagc 420 gagttccgca tggtttgtcg tacgcaatta gctgtttatt gttttttttt tggggggggg 480 aagagaaaaa gaaaaaagga g 501 <210> SEQ ID NO 14 <211> LENGTH: 106 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 14 Gly Arg Val Tyr Glu Glu Ile Pro Trp Val Arg Val Tyr Glu Asn Ile 1 5 10 15 Cys Leu Arg Arg Gln Asp Ala Gly Gly Ala Ala Pro Pro Gly Asp Ala 20 25 30 Pro Asp Ser Pro Tyr Ile Glu Ala Glu Asn Pro Leu Tyr Asp Trp Gly 35 40 45 Gly Ser Ala Leu Phe Ser Pro Pro Gly Ala Thr Arg Ala Pro Asp Pro 50 55 60 Gly Leu Ser Leu Ser Pro Met Pro Ala Arg Pro Arg Thr Asn Ala Leu 65 70 75 80 Ala Asn Asp Gly Pro Thr Asn Val Ala Ala Leu Ser Ala Leu Leu Thr 85 90 95 Lys Leu Lys Arg Gly Arg His Gln Ser His 100 105 <210> SEQ ID NO 15 <211> LENGTH: 722 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 15 Met Gln Arg Arg Ala Arg Gly Ala Ser Ser Leu Arg Leu Ala Arg Cys 1 5 10 15 Leu Thr Pro Ala Asn Leu Ile Arg Gly Ala Asn Ala Gly Val Pro Glu 20 25 30 Arg Arg Ile Phe Ala Gly Cys Leu Leu Pro Thr Pro Glu Gly Leu Leu 35 40 45 Ser Ala Ala Val Gly Val Leu Arg Gln Arg Ala Asp Asp Leu Gln Pro 50 55 60 Ala Phe Leu Thr Gly Ala Asp Arg Ser Val Arg Leu Ala Ala Arg His 65 70 75 80 His Asn Thr Val Pro Glu Ser Leu Ile Val Asp Gly Leu Ala Ser Asp 85 90 95 Pro His Tyr Asp Tyr Ile Arg His Tyr Ala Ser Ala Ala Lys Gln Ala 100 105 110 Leu Gly Glu Val Glu Leu Ser Gly Gly Gln Leu Ser Arg Ala Ile Leu 115 120 125 Ala Gln Tyr Trp Lys Tyr Leu Gln Thr Val Val Pro Ser Gly Leu Asp 130 135 140 Ile Pro Asp Asp Pro Ala Gly Asp Cys Asp Pro Ser Leu His Val Leu 145 150 155 160 Leu Arg Pro Thr Leu Leu Pro Lys Leu Leu Val Arg Ala Pro Phe Lys 165 170 175 Ser Gly Ala Ala Ala Ala Lys Tyr Ala Ala Ala Val Ala Gly Leu Arg 180 185 190 Asp Ala Ala His Arg Leu Gln Gln Tyr Met Phe Phe Met Arg Pro Ala 195 200 205 Asp Pro Ser Arg Pro Ser Thr Asp Thr Ala Leu Arg Leu Ser Glu Leu 210 215 220 Leu Ala Tyr Val Ser Val Leu Tyr His Trp Ala Ser Trp Met Leu Trp 225 230 235 240 Thr Ala Asp Lys Tyr Val Cys Arg Arg Leu Gly Pro Ala Asp Arg Arg 245 250 255 Phe Val Ala Leu Ser Gly Ser Leu Glu Ala Pro Ala Glu Thr Phe Ala 260 265 270 Arg His Leu Asp Arg Gly Pro Ser Gly Thr Thr Gly Ser Met Gln Cys 275 280 285 Met Ala Leu Arg Ala Ala Val Ser Asp Val Leu Gly His Leu Thr Arg 290 295 300 Leu Ala His Leu Trp Glu Thr Gly Lys Arg Ser Gly Gly Thr Tyr Gly 305 310 315 320 Ile Val Asp Ala Ile Val Ser Thr Val Glu Val Leu Ser Ile Val His 325 330 335 His His Ala Gln Tyr Ile Ile Asn Ala Thr Leu Thr Gly Tyr Val Val 340 345 350 Trp Ala Ser Asp Ser Leu Asn Asn Glu Tyr Leu Thr Ala Ala Val Asp 355 360 365 Ser Gln Glu Arg Phe Cys Arg Thr Ala Ala Pro Leu Phe Pro Thr Met 370 375 380 Thr Ala Pro Ser Trp Ala Arg Met Glu Leu Ser Ile Lys Ser Trp Phe 385 390 395 400 Gly Ala Ala Leu Ala Pro Asp Leu Leu Arg Ser Gly Thr Pro Ser Pro 405 410 415 His Tyr Glu Ser Ile Leu Arg Leu Ala Ala Ser Gly Pro Pro Gly Gly 420 425 430 Arg Gly Ala Val Gly Gly Ser Cys Arg Asp Lys Ile Gln Arg Thr Arg 435 440 445 Arg Asp Asn Ala Pro Pro Pro Leu Pro Arg Ala Arg Pro His Ser Thr 450 455 460 Pro Ala Ala Pro Arg Arg Cys Arg Arg His Arg Glu Asp Leu Pro Glu 465 470 475 480 Pro Pro His Val Asp Ala Ala Asp Arg Gly Pro Glu Pro Cys Ala Gly 485 490 495 Arg Pro Ala Thr Tyr Tyr Thr His Met Ala Gly Ala Pro Pro Arg Leu 500 505 510 Pro Pro Arg Asn Pro Ala Pro Pro Glu Gln Arg Pro Ala Ala Ala Ala 515 520 525 Arg Pro Leu Ala Ala Gln Arg Glu Ala Ala Gly Val Tyr Asp Ala Val 530 535 540 Arg Thr Trp Gly Pro Asp Ala Glu Ala Glu Pro Asp Gln Met Glu Asn 545 550 555 560 Thr Tyr Leu Leu Pro Asp Asp Asp Ala Ala Met Pro Ala Gly Val Gly 565 570 575 Leu Gly Ala Thr Pro Ala Ala Asp Thr Thr Ala Ala Ala Ala Trp Pro 580 585 590 Ala Glu Ser His Ala Pro Arg Ala Pro Ser Glu Asp Ala Asp Ser Ile 595 600 605 Tyr Glu Ser Val Gly Glu Asp Gly Gly Arg Val Tyr Glu Glu Ile Pro 610 615 620 Trp Val Arg Val Tyr Glu Asn Ile Cys Pro Arg Arg Arg Leu Ala Gly 625 630 635 640 Gly Ala Ala Leu Pro Gly Asp Ala Pro Asp Ser Pro Tyr Ile Glu Ala 645 650 655 Glu Asn Pro Leu Tyr Asp Trp Gly Gly Ser Ala Leu Phe Ser Pro Arg 660 665 670 Arg Ala Thr Arg Ala Pro Asp Pro Gly Leu Ser Leu Ser Pro Met Pro 675 680 685 Ala Arg Pro Arg Thr Asn Ala Leu Ala Asn Asp Gly Pro Thr Asn Val 690 695 700 Ala Ala Leu Ser Ala Leu Leu Thr Lys Leu Lys Arg Gly Arg His Gln 705 710 715 720 Ser His <210> SEQ ID NO 16 <211> LENGTH: 200 <212> TYPE: DNA <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 16 actgcaacgc aatcccatga aggccctgta tccgctcacc accaaggaac tcaagacttc 60 cgaccccggg ggcgtgggcg gggaggggga ggaaggcgcg gaggggggcg ggtttgacga 120 ggccaagttg gccgaggccc gagaaatgat ccgatatatg gctttggtgt cggccatgga 180 gcgcacggaa cacaaggcca 200 <210> SEQ ID NO 17 <211> LENGTH: 66 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 17 Leu Gln Arg Asn Pro Met Lys Ala Leu Tyr Pro Leu Thr Thr Lys Glu 1 5 10 15 Leu Lys Thr Ser Asp Pro Gly Gly Val Gly Gly Glu Gly Glu Glu Gly 20 25 30 Ala Glu Gly Gly Gly Phe Asp Glu Ala Lys Leu Ala Glu Ala Arg Glu 35 40 45 Met Ile Arg Tyr Met Ala Leu Val Ser Ala Met Glu Arg Thr Glu His 50 55 60 Lys Ala 65 <210> SEQ ID NO 18 <211> LENGTH: 904 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 18 Met Arg Gly Gly Gly Leu Ile Cys Ala Leu Val Val Gly Ala Leu Val 1 5 10 15 Ala Ala Val Ala Ser Ala Ala Pro Ala Ala Pro Ala Ala Pro Arg Ala 20 25 30 Ser Gly Gly Val Ala Ala Thr Val Ala Ala Asn Gly Gly Pro Ala Ser 35 40 45 Arg Pro Pro Pro Val Pro Ser Pro Ala Thr Thr Lys Ala Arg Lys Arg 50 55 60 Lys Thr Lys Lys Pro Pro Lys Arg Pro Glu Ala Thr Pro Pro Pro Asp 65 70 75 80 Ala Asn Ala Thr Val Ala Ala Gly His Ala Thr Leu Arg Ala His Leu 85 90 95 Arg Glu Ile Lys Val Glu Asn Ala Asp Ala Gln Phe Tyr Val Cys Pro 100 105 110 Pro Pro Thr Gly Ala Thr Val Val Gln Phe Glu Gln Pro Arg Arg Cys 115 120 125 Pro Thr Arg Pro Glu Gly Gln Asn Tyr Thr Glu Gly Ile Ala Val Val 130 135 140 Phe Lys Glu Asn Ile Ala Pro Tyr Lys Phe Lys Ala Thr Met Tyr Tyr 145 150 155 160 Lys Asp Val Thr Val Ser Gln Val Trp Phe Gly His Arg Tyr Ser Gln 165 170 175 Phe Met Gly Ile Phe Glu Asp Arg Ala Pro Val Pro Phe Glu Glu Val 180 185 190 Ile Asp Lys Ile Asn Thr Lys Gly Val Cys Arg Ser Thr Ala Lys Tyr 195 200 205 Val Arg Asn Asn Met Glu Thr Thr Ala Phe His Arg Asp Asp His Glu 210 215 220 Thr Asp Met Glu Leu Lys Pro Ala Lys Val Ala Thr Arg Thr Ser Arg 225 230 235 240 Gly Trp His Thr Thr Asp Leu Lys Tyr Asn Pro Ser Arg Val Glu Ala 245 250 255 Phe His Arg Tyr Gly Thr Thr Val Asn Cys Ile Val Glu Glu Val Asp 260 265 270 Ala Arg Ser Val Tyr Pro Tyr Asp Glu Phe Val Leu Ala Thr Gly Asp 275 280 285 Phe Val Tyr Met Ser Pro Phe Tyr Gly Tyr Arg Glu Gly Ser His Thr 290 295 300 Glu His Thr Ser Tyr Ala Ala Asp Arg Phe Lys Gln Val Asp Gly Phe 305 310 315 320 Tyr Ala Arg Asp Leu Thr Thr Lys Ala Arg Ala Thr Ser Pro Thr Thr 325 330 335 Arg Asn Leu Leu Thr Thr Pro Lys Phe Thr Val Ala Trp Asp Trp Val 340 345 350 Pro Lys Arg Pro Ala Val Cys Thr Met Thr Lys Trp Gln Glu Val Asp 355 360 365 Glu Met Leu Arg Ala Glu Tyr Gly Gly Ser Phe Arg Phe Ser Ser Asp 370 375 380 Ala Ile Ser Thr Thr Phe Thr Thr Asn Leu Thr Glu Tyr Ser Leu Ser 385 390 395 400 Arg Val Asp Leu Gly Asp Cys Ile Gly Arg Asp Ala Arg Glu Ala Ile 405 410 415 Asp Arg Met Phe Ala Arg Lys Tyr Asn Ala Thr His Ile Lys Val Gly 420 425 430 Gln Pro Gln Tyr Tyr Leu Ala Thr Gly Gly Phe Leu Ile Ala Tyr Gln 435 440 445 Pro Leu Leu Ser Asn Thr Leu Ala Glu Leu Tyr Val Arg Glu Tyr Met 450 455 460 Arg Glu Gln Asp Arg Lys Pro Arg Asn Ala Thr Pro Ala Pro Leu Arg 465 470 475 480 Glu Ala Pro Ser Ala Asn Ala Ser Val Glu Arg Ile Lys Thr Thr Ser 485 490 495 Ser Ile Glu Phe Ala Arg Leu Gln Phe Thr Tyr Asn His Ile Gln Arg 500 505 510 His Val Asn Asp Met Leu Gly Arg Ile Ala Val Ala Trp Cys Glu Leu 515 520 525 Gln Asn His Glu Leu Thr Leu Trp Asn Glu Ala Arg Lys Leu Asn Pro 530 535 540 Asn Ala Ile Ala Ser Ala Thr Val Gly Arg Arg Val Ser Ala Arg Met 545 550 555 560 Leu Gly Asp Val Met Ala Val Ser Thr Cys Val Pro Val Ala Pro Asp 565 570 575 Asn Val Ile Val Gln Asn Ser Met Arg Val Ser Ser Arg Pro Gly Thr 580 585 590 Cys Tyr Ser Arg Pro Leu Val Ser Phe Arg Tyr Glu Asp Gln Gly Pro 595 600 605 Leu Ile Glu Gly Gln Leu Gly Glu Asn Asn Glu Leu Arg Leu Thr Arg 610 615 620 Asp Ala Leu Glu Pro Cys Thr Val Gly His Arg Arg Tyr Phe Ile Phe 625 630 635 640 Gly Gly Gly Tyr Val Tyr Phe Glu Glu Tyr Ala Tyr Ser His Gln Leu 645 650 655 Ser Arg Ala Asp Val Thr Thr Val Ser Thr Phe Ile Asp Leu Asn Ile 660 665 670 Thr Met Leu Glu Asp His Glu Phe Val Pro Leu Glu Val Tyr Thr Arg 675 680 685 His Glu Ile Lys Asp Ser Gly Leu Leu Asp Tyr Thr Glu Val Gln Arg 690 695 700 Arg Asn Gln Leu His Asp Leu Arg Phe Ala Asp Ile Asp Thr Val Ile 705 710 715 720 Arg Ala Asp Ala Asn Ala Ala Met Phe Ala Gly Leu Cys Ala Phe Phe 725 730 735 Glu Gly Met Gly Asp Leu Gly Arg Ala Val Gly Lys Val Val Met Gly 740 745 750 Val Val Gly Gly Val Val Ser Ala Val Ser Gly Val Ser Ser Phe Met 755 760 765 Ser Asn Pro Phe Gly Ala Leu Ala Val Gly Leu Leu Val Leu Ala Gly 770 775 780 Leu Val Ala Ala Phe Phe Ala Phe Arg Tyr Val Leu Gln Leu Gln Arg 785 790 795 800 Asn Pro Met Lys Ala Leu Tyr Pro Leu Thr Thr Lys Glu Leu Lys Thr 805 810 815 Ser Asp Pro Gly Gly Val Gly Gly Glu Gly Glu Glu Gly Ala Glu Gly 820 825 830 Gly Gly Phe Asp Glu Ala Lys Leu Ala Glu Ala Arg Glu Met Ile Arg 835 840 845 Tyr Met Ala Leu Val Ser Ala Met Glu Arg Thr Glu His Lys Ala Arg 850 855 860 Lys Lys Gly Thr Ser Ala Leu Leu Ser Ser Lys Val Thr Asn Met Val 865 870 875 880 Leu Arg Lys Arg Asn Lys Ala Arg Tyr Ser Pro Leu His Asn Glu Asp 885 890 895 Glu Ala Gly Asp Glu Asp Glu Leu 900 <210> SEQ ID NO 19 <211> LENGTH: 443 <212> TYPE: DNA <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 19 ccctctccca cacggtcggt gccccccatc tctgtttcat catcgtcccg gttgcgttgc 60 gctttccggc cctcccgcac ccccgcgttc cggtgtctcg cggcccggcg ccatgatcac 120 ggattgtttc gaagcagaca tcgcgatccc ctcgggtatc tcgcgccccg atgccgcggc 180 gctgcagcgg tgcgagggtc gagtggtctt tctgccgacc atccgccgcc agctggcgct 240 cgcggacgtg gcgcacgaat cgttcgtctc cggaggagtt agtcccgaca cgttggggtt 300 gttgctggcg taccgcaggc gcttccccgc ggtaatcacg cgggtgctgc ccacgcgaat 360 cgtcgcctgc cccgtggacc tggggctcac gcacgccggc accgtcaatc tccgcaacac 420 ctcccccgtc gacctctgca acg 443 <210> SEQ ID NO 20 <211> LENGTH: 37 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 20 Pro Leu Pro His Gly Arg Cys Pro Pro Ser Leu Phe His His Arg Pro 1 5 10 15 Gly Cys Val Ala Leu Ser Gly Pro Pro Ala Pro Pro Arg Ser Gly Val 20 25 30 Ser Arg Pro Gly Ala 35 <210> SEQ ID NO 21 <211> LENGTH: 147 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 21 Pro Leu Pro His Gly Arg Cys Pro Pro Ser Leu Phe His His Arg Pro 1 5 10 15 Gly Cys Val Ala Leu Ser Gly Pro Pro Ala Pro Pro Arg Ser Gly Val 20 25 30 Ser Arg Pro Gly Ala Met Ile Thr Asp Cys Phe Glu Ala Asp Ile Ala 35 40 45 Ile Pro Ser Gly Ile Ser Arg Pro Asp Ala Ala Ala Leu Gln Arg Cys 50 55 60 Glu Gly Arg Val Val Phe Leu Pro Thr Ile Arg Arg Gln Leu Ala Leu 65 70 75 80 Ala Asp Val Ala His Glu Ser Phe Val Ser Gly Gly Val Ser Pro Asp 85 90 95 Thr Leu Gly Leu Leu Leu Ala Tyr Arg Arg Arg Phe Pro Ala Val Ile 100 105 110 Thr Arg Val Leu Pro Thr Arg Ile Val Ala Cys Pro Val Asp Leu Gly 115 120 125 Leu Thr His Ala Gly Thr Val Asn Leu Arg Asn Thr Ser Pro Val Asp 130 135 140 Leu Cys Asn 145 <210> SEQ ID NO 22 <211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 22 Met Ile Thr Asp Cys Phe Glu Ala Asp Ile Ala Ile Pro Ser Gly Ile 1 5 10 15 Ser Arg Pro Asp Ala Ala Ala Leu Gln Arg Cys Glu Gly Arg Val Val 20 25 30 Phe Leu Pro Thr Ile Arg Arg Gln Leu Ala Leu Ala Asp Val Ala His 35 40 45 Glu Ser Phe Val Ser Gly Gly Val Ser Pro Asp Thr Leu Gly Leu Leu 50 55 60 Leu Ala Tyr Arg Arg Arg Phe Pro Ala Val Ile Thr Arg Val Leu Pro 65 70 75 80 Thr Arg Ile Val Ala Cys Pro Val Asp Leu Gly Leu Thr His Ala Gly 85 90 95 Thr Val Asn Leu Arg Asn Thr Ser Pro Val Asp Leu Cys Asn 100 105 110 <210> SEQ ID NO 23 <211> LENGTH: 318 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 23 Met Ile Thr Asp Cys Phe Glu Ala Asp Ile Ala Ile Pro Ser Gly Ile 1 5 10 15 Ser Arg Pro Asp Ala Ala Ala Leu Gln Arg Cys Glu Gly Arg Val Val 20 25 30 Phe Leu Pro Thr Ile Arg Arg Gln Leu Ala Leu Ala Asp Val Ala His 35 40 45 Glu Ser Phe Val Ser Gly Gly Val Ser Pro Asp Thr Leu Gly Leu Leu 50 55 60 Leu Ala Tyr Arg Arg Arg Phe Pro Ala Val Ile Thr Arg Val Leu Pro 65 70 75 80 Thr Arg Ile Val Ala Cys Pro Val Asp Leu Gly Leu Thr His Ala Gly 85 90 95 Thr Val Asn Leu Arg Asn Thr Ser Pro Val Asp Leu Cys Asn Gly Asp 100 105 110 Pro Val Ser Leu Val Pro Pro Val Phe Glu Gly Gln Ala Thr Asp Val 115 120 125 Arg Leu Glu Ser Leu Asp Leu Thr Leu Arg Phe Pro Val Pro Leu Pro 130 135 140 Thr Pro Leu Ala Arg Glu Ile Val Ala Arg Leu Val Ala Arg Gly Ile 145 150 155 160 Arg Asp Leu Asn Pro Asp Pro Arg Thr Pro Gly Glu Leu Pro Asp Leu 165 170 175 Asn Val Leu Tyr Tyr Asn Gly Ala Arg Leu Ser Leu Val Ala Asp Val 180 185 190 Gln Gln Leu Ala Ser Val Asn Thr Glu Leu Arg Ser Leu Val Leu Asn 195 200 205 Met Val Tyr Ser Ile Thr Glu Gly Thr Thr Leu Ile Leu Thr Leu Ile 210 215 220 Pro Arg Leu Leu Ala Leu Ser Ala Gln Asp Gly Tyr Val Asn Ala Leu 225 230 235 240 Leu Gln Met Gln Ser Val Thr Arg Glu Ala Ala Gln Leu Ile His Pro 245 250 255 Glu Ala Pro Met Leu Met Gln Asp Gly Glu Arg Arg Leu Pro Leu Tyr 260 265 270 Glu Ala Leu Val Ala Trp Leu Ala His Ala Gly Gln Leu Gly Asp Ile 275 280 285 Leu Ala Leu Ala Pro Ala Val Arg Val Cys Thr Phe Asp Gly Ala Ala 290 295 300 Val Val Gln Ser Gly Asp Met Ala Pro Val Ile Arg Tyr Pro 305 310 315 <210> SEQ ID NO 24 <211> LENGTH: 502 <212> TYPE: DNA <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 24 actgttgtag gggggaaaac acagttccgg gaaggcgttt attgcggaga gaggggggaa 60 agaaagagaa acaaaagaaa cggcaagaaa gactcaagac gtgcgcgtga tcggaaaaaa 120 ggccgggggg atcccggtcg gggccgccag gtaaatggcc atgatgaccg cgaccatgag 180 gtcgtccgcg gcaccgttgc gttttccgga gtacatgcgg acgtcggtgt tgggagagac 240 ggtttcgatg aggttgttga gctgctcgga cagatactcg accgggtcgg tctgcaggcg 300 caccgtcacg gagacgagct cctgggacgc catgacgccc ccggagttga actttttgat 360 aaagtattcg aaggcgggcg tcttctgttt gttgagcaga aagaaggggt acaataccgc 420 gccgccgggc ggctcgcagt gatagaagag gagctcgggc cccgggccgt tggcccccgc 480 cgaggccagg atgcggtgca tc 502 <210> SEQ ID NO 25 <211> LENGTH: 135 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 25 Met His Arg Ile Leu Ala Ser Ala Gly Ala Asn Gly Pro Gly Pro Glu 1 5 10 15 Leu Leu Phe Tyr His Cys Glu Pro Pro Gly Gly Ala Val Leu Tyr Pro 20 25 30 Phe Phe Leu Leu Asn Lys Gln Lys Thr Pro Ala Phe Glu Tyr Phe Ile 35 40 45 Lys Lys Phe Asn Ser Gly Gly Val Met Ala Ser Gln Glu Leu Val Ser 50 55 60 Val Thr Val Arg Leu Gln Thr Asp Pro Val Glu Tyr Leu Ser Glu Gln 65 70 75 80 Leu Asn Asn Leu Ile Glu Thr Val Ser Pro Asn Thr Asp Val Arg Met 85 90 95 Tyr Ser Gly Lys Arg Asn Gly Ala Ala Asp Asp Leu Met Val Ala Val 100 105 110 Ile Met Ala Ile Tyr Leu Ala Ala Pro Thr Gly Ile Pro Pro Ala Phe 115 120 125 Phe Pro Ile Thr Arg Thr Ser 130 135 <210> SEQ ID NO 26 <211> LENGTH: 734 <212> TYPE: PRT <213> ORGANISM: Herpes simplex virus <400> SEQUENCE: 26 Met Phe Gly Gln Gln Leu Ala Ser Asp Val Gln Gln Tyr Leu Glu Arg 1 5 10 15 Leu Glu Lys Gln Arg Gln Gln Lys Val Gly Val Asp Glu Ala Ser Ala 20 25 30 Gly Leu Thr Leu Gly Gly Asp Ala Leu Arg Val Pro Phe Leu Asp Phe 35 40 45 Ala Thr Ala Thr Pro Lys Arg His Gln Thr Val Val Pro Gly Val Gly 50 55 60 Thr Leu His Asp Cys Cys Glu His Ser Pro Leu Phe Ser Ala Val Ala 65 70 75 80 Arg Arg Leu Leu Phe Asn Ser Leu Val Pro Ala Gln Leu Arg Gly Arg 85 90 95 Asp Phe Gly Gly Asp His Thr Ala Lys Leu Glu Phe Leu Ala Pro Glu 100 105 110 Leu Val Arg Ala Val Ala Arg Leu Arg Phe Arg Glu Cys Ala Pro Glu 115 120 125 Asp Ala Val Pro Gln Arg Asn Ala Tyr Tyr Ser Val Leu Asn Thr Phe 130 135 140 Gln Ala Leu His Arg Ser Glu Ala Phe Arg Gln Leu Val His Phe Val 145 150 155 160 Arg Asp Phe Ala Gln Leu Leu Lys Thr Ser Phe Arg Ala Ser Ser Leu 165 170 175 Ala Glu Thr Thr Gly Pro Pro Lys Lys Arg Ala Lys Val Asp Val Ala 180 185 190 Thr His Gly Gln Thr Tyr Gly Thr Leu Glu Leu Phe Gln Lys Met Ile 195 200 205 Leu Met His Ala Thr Tyr Phe Leu Ala Ala Val Leu Leu Gly Asp His 210 215 220 Ala Glu Gln Val Asn Thr Phe Leu Arg Leu Val Phe Glu Ile Pro Leu 225 230 235 240 Phe Ser Asp Thr Ala Val Arg His Phe Arg Gln Arg Ala Thr Val Phe 245 250 255 Leu Val Pro Arg Arg His Gly Lys Thr Trp Phe Leu Val Pro Leu Ile 260 265 270 Ala Leu Ser Leu Ala Ser Phe Arg Gly Ile Lys Ile Gly Tyr Thr Ala 275 280 285 His Ile Arg Lys Ala Thr Glu Pro Val Phe Asp Glu Ile Asp Ala Cys 290 295 300 Leu Arg Gly Trp Phe Gly Ser Ser Arg Val Asp His Val Lys Gly Glu 305 310 315 320 Thr Ile Ser Phe Ser Phe Pro Asp Gly Ser Arg Ser Thr Ile Val Phe 325 330 335 Ala Ser Ser His Asn Thr Asn Gly Ile Arg Gly Gln Asp Phe Asn Leu 340 345 350 Leu Phe Val Asp Glu Ala Asn Phe Ile Arg Pro Asp Ala Val Gln Thr 355 360 365 Ile Met Gly Phe Leu Asn Gln Ala Asn Cys Lys Ile Ile Phe Val Ser 370 375 380 Ser Thr Asn Thr Gly Lys Ala Ser Thr Ser Phe Leu Tyr Asn Leu Arg 385 390 395 400 Gly Ala Ala Asp Glu Leu Leu Asn Val Val Thr Tyr Ile Cys Asp Asp 405 410 415 His Met Pro Arg Val Val Thr His Thr Asn Ala Thr Ala Cys Ser Cys 420 425 430 Tyr Ile Leu Asn Lys Pro Val Phe Ile Thr Met Asp Gly Ala Val Arg 435 440 445 Arg Thr Ala Asp Leu Phe Leu Pro Asp Ser Phe Met Gln Glu Ile Ile 450 455 460 Gly Gly Gln Ala Arg Glu Thr Gly Asp Asp Arg Pro Val Leu Thr Lys 465 470 475 480 Ser Ala Gly Glu Arg Phe Leu Leu Tyr Arg Pro Ser Thr Thr Thr Asn 485 490 495 Ser Gly Leu Met Ala Pro Glu Leu Tyr Val Tyr Val Asp Pro Ala Phe 500 505 510 Thr Ala Asn Thr Arg Ala Ser Gly Thr Gly Ile Ala Val Val Gly Arg 515 520 525 Tyr Arg Asp Asp Phe Ile Ile Phe Ala Leu Glu His Phe Phe Leu Arg 530 535 540 Ala Leu Thr Gly Ser Ala Pro Ala Asp Ile Ala Arg Cys Val Val His 545 550 555 560 Ser Leu Ala Gln Val Leu Ala Leu His Pro Gly Ala Phe Arg Ser Val 565 570 575 Arg Val Ala Val Glu Gly Asn Ser Ser Gln Asp Ser Ala Val Ala Ile 580 585 590 Ala Thr His Val His Thr Glu Met His Arg Ile Leu Ala Ser Ala Gly 595 600 605 Ala Asn Gly Pro Gly Pro Glu Leu Leu Phe Tyr His Cys Glu Pro Pro 610 615 620 Gly Gly Ala Val Leu Tyr Pro Phe Phe Leu Leu Asn Lys Gln Lys Thr 625 630 635 640 Pro Ala Phe Glu Tyr Phe Ile Lys Lys Phe Asn Ser Gly Gly Val Met 645 650 655 Ala Ser Gln Glu Leu Val Ser Val Thr Val Arg Leu Gln Thr Asp Pro 660 665 670 Val Glu Tyr Leu Ser Glu Gln Leu Asn Asn Leu Ile Glu Thr Val Ser 675 680 685 Pro Asn Thr Asp Val Arg Met Tyr Ser Gly Lys Arg Asn Gly Ala Ala 690 695 700 Asp Asp Leu Met Val Ala Val Ile Met Ala Ile Tyr Leu Ala Ala Pro 705 710 715 720 Thr Gly Ile Pro Pro Ala Phe Phe Pro Ile Thr Arg Thr Ser 725 730 <210> SEQ ID NO 27 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 27 Gly Arg Val Tyr Glu Glu Ile Pro Trp Val Arg Val Tyr Glu Asn 5 10 15 <210> SEQ ID NO 28 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 28 Tyr Glu Asn Ile Cys Leu Arg Arg Gln Asp Ala Gly Gly Ala Ala 5 10 15 <210> SEQ ID NO 29 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 29 Pro Asp Ser Pro Tyr Ile Glu Ala Glu Asn Pro Leu Tyr Asp Trp 5 10 15 <210> SEQ ID NO 30 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 30 Tyr Ile Glu Ala Glu Asn Pro Leu Tyr Asp Trp Gly Gly Ser Ala 5 10 15 <210> SEQ ID NO 31 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 31 Thr Asn Ala Leu Ala Asn Asp Gly Pro Thr Asn Val Ala Ala Leu 5 10 15 <210> SEQ ID NO 32 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 32 Arg Val Leu Pro Thr Arg Ile Val Ala Cys Pro Val Asp Leu Gly 5 10 15 <210> SEQ ID NO 33 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 33 Thr Arg Ile Val Ala Cys Pro Val Asp Leu Gly Leu Thr His Ala 5 10 15 <210> SEQ ID NO 34 <211> LENGTH: 661 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 34 ctcctcttcc gcctcctcct cctcctcttc cgcctcctcc tcctcctcct ccgcctcttc 60 ctctgcgggc ggggctggtg ggagcgtcgc gtccgcgtcc ggcgctgggg agagacgaga 120 aacctccctc ggcccccgcg ctgctgcgcc gcgggggccg aggaagtgtg ccaggaagac 180 gcgccacgcg gagggcggcc ccgagcccgg ggcccgcgac ccggcgcccg gcctcacgcg 240 ctacctgccc atcgcggggg tctcgagcgt cgtggccctg gcgccttacg tgaacaagac 300 ggtcacgggg gactgcctgc ccgtcctgga catggagacg ggccacatag gggcctacgt 360 ggtcctcgtg gaccagacgg ggaacgtggc ggacctgctg cgggccgcgg cccccgcgtg 420 gagccgccgc accctgctcc ccgagcacgc gcgcaactgc gtgaggcccc ccgactaccc 480 gacgcccccc gcgtcggagt ggaacagcct ctggatgacc ccggtgggca acatgctctt 540 tgaccagggc accctggtgg gcgcgctgga cttccacggc ctccggtcgc gccacccgtg 600 gtctcgggag cagggcgcgc ccgcgccggc cggcgacgcc cccgcgggcc acggggagta 660 g 661 <210> SEQ ID NO 35 <211> LENGTH: 2481 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 35 atggaacccc ggcccggcac gagctcccgg gcggaccccg gccccgagcg gccgccgcgg 60 cagacccccg gcacgcagcc cgccgccccg cacgcctggg ggatgctcaa cgacatgcag 120 tggctcgcca gcagcgactc ggaggaggag accgaggtgg gaatctctga cgacgacctt 180 caccgcgact ccacctccga ggcgggcagc acggacacgg agatgttcga ggcgggcctg 240 atggacgcgg ccacgccccc ggcccggccc ccggccgagc gccagggcag ccccacgccc 300 gccgacgcgc agggatcctg tgggggtggg cccgtgggtg aggaggaagc ggaagcggga 360 ggggggggcg acgtgtgtgc cgtgtgcacg gacgagatcg ccccgcccct gcgctgccag 420 agttttccct gcctgcaccc cttctgcatc ccgtgcatga agacctggat tccgttgcgc 480 aacacgtgtc ccctgtgcaa caccccggtg gcgtacctga tagtgggcgt gaccgccagc 540 gggtcgttca gcaccatccc gatagtgaac gacccccgga cccgcgtgga ggccgaggcg 600 gccgtgcggg ccggcacggc cgtggacttt atctggacgg gcaacccgcg gacggccccg 660 cgctccctgt cgctgggggg acacacggtc cgcgccctgt cgcccacccc cccgtggccc 720 ggcacggacg acgaggacga tgacctggcc gacggtgtgg actacgtccc gcccgccccc 780 cgaagagcgc cccggcgcgg gggcggcggt gcgggggcga cccgcggaac ctcccagccc 840 gccgcgaccc gaccggcgcc ccctggcgcc ccgcggagca gcagcagcgg cggcgccccg 900 ttgcgggcgg gggtgggatc tgggtctggg ggcggccctg ccgtcgcggc cgtcgtgccg 960 agagtggcct ctcttccccc tgcggccggc ggggggcgcg cgcaggcgcg gcgggtgggc 1020 gaagacgccg cggcggcgga gggcaggacg ccccccgcga gacagccccg cgcggcccag 1080 gagcccccca tagtcatcag cgactctccc ccgccgtctc cgcgccgccc cgcgggcccc 1140 gggccgctct cctttgtctc ctcctcctcc gcacaggtgt cctcgggccc cgggggggga 1200 ggtctgccac agtcgtcggg gcgcgccgcg cgcccccgcg cggccgtcgc cccgcgcgtc 1260 cggagtccgc cccgcgccgc cgccgccccc gtggtgtctg cgagcgcgga cgcggccggg 1320 cccgcgccgc ccgccgtgcc ggtggacgcg caccgcgcgc cccggtcgcg catgacccag 1380 gctcagaccg acacccaagc acagagtctg ggccgggcag gcgcgaccga cgcgcgcggg 1440 tcgggagggc cgggcgcgga gggaggaccc ggggtccccc gcggcaccaa cacccccggt 1500 gccgcccccc acgccgcgga gggggcggcg gcccgccccc ggaagaggcg cgggtcggac 1560 tcgggccccg cggcctcgtc ctccgcctct tcctccgccg ccccgcgctc gcccctcgcc 1620 ccccaggggg tgggggccaa gagggcggcg ccgcgccggg ccccggactc ggactcgggc 1680 gaccgcggcc acgggccgct cgccccggcg tccgcgggcg ccgcgccccc gtcggcgtct 1740 ccgtcgtccc aggccgcggt cgccgccgcc tcctcctcct ccgcctcctc ctcctccgcc 1800 tcctcctcct ccgcctcctc ctcctccgcc tcctcctcct ccgcctcctc ctcctccgcc 1860 tcctcctcct ccgcctcttc ctctgcgggc ggggctggtg ggagcgtcgc gtccgcgtcc 1920 ggcgctgggg agagacgaga aacctccctc ggcccccgcg ctgctgcgcc gcgggggccg 1980 aggaagtgtg ccaggaagac gcgccacgcg gagggcggcc ccgagcccgg ggcccgcgac 2040 ccggcgcccg gcctcacgcg ctacctgccc atcgcggggg tctcgagcgt cgtggccctg 2100 gcgccttacg tgaacaagac ggtcacgggg gactgcctgc ccgtcctgga catggagacg 2160 ggccacatag gggcctacgt ggtcctcgtg gaccagacgg ggaacgtggc ggacctgctg 2220 cgggccgcgg cccccgcgtg gagccgccgc accctgctcc ccgagcacgc gcgcaactgc 2280 gtgaggcccc ccgactaccc gacgcccccc gcgtcggagt ggaacagcct ctggatgacc 2340 ccggtgggca acatgctctt tgaccagggc accctggtgg gcgcgctgga cttccacggc 2400 ctccggtcgc gccacccgtg gtctcgggag cagggcgcgc ccgcgccggc cggcgacgcc 2460 cccgcgggcc acggggagta g 2481 <210> SEQ ID NO 36 <211> LENGTH: 1603 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 36 cggccggagg gctgtcccgc atcgatatca cgagccccat gaagcccttc ccgtatcgcg 60 cgcgcacgag cgcggcgtcg cacccgaacg ccagcccgcc cgtcgtccag acgcccacgg 120 gccacgtcga ggccgacggg gagaggtaca cgtaccgacc cggagtccgt agcaggcccc 180 tggcggccag ccaggtcacg gatgcgttgt gcagatgcgc gatgctcagg ttcgtcgtcg 240 gatgcctcgg tgtccccgcg ggcggccccg ggggcggcgc gttgcgtcgg ccgtccgggt 300 gcctctcggt cgccccgtcg tctccccgcg ggaacgtaag cccctcgcgg tccgcgcggc 360 cgcgaatgtt acccaggccc gggaccgcaa cagcgcggag gcgccggggt tgtgcgacag 420 tcccttgagc tgggtcacct cggcgggggg acgggacgtg ggccccgcct cggggagctc 480 gggcaggctc gcgttccgag gccggccgag cagataggtc tttgggatgt aaagcagctg 540 cccggggtcc cgaggaaact cggccgtggt gaccaacacg aaacaaaagc gctcggcgta 600 ccaccgaagc atgggcacgg atgccgtagt caggttgagt tcgcccgggg gcgccaagcg 660 tccgcgctgg gggtcgctgg cgtcgggggt tgttgggcaa ccacagacgc ccggtgtttt 720 gtcgcgccag tacgtgcggg ccaaccccag accgtgcaaa aaccacgggt cgatttgctc 780 cgtccagtac gtgtcatggc ccccggcaac gcccaccagg acccccatca ccacccacag 840 accggggccc atggtcgtcc gtcccggctg ccagtccgca gatggggggg ggtgtccgta 900 cccacggccc aaagaggctc cgcacctcgg aggctatcgg aggccctttg ttgccgtaag 960 cgcgggccaa aggatggggt ggggtgaggg taaaagcaca aagggagtac cagaccgaaa 1020 acaaggacgg atcggcccgc tccgtttttc ggtggggtgc tgatacggtg ccagccctgg 1080 ccccgaaccc ccgcgcttat ggacacacca cacgacaaca atgcctttta ttctgttctt 1140 ttattgccgt catcgccggg aggccttccg ttcgggcttc cgtgtttgaa ctaaactccc 1200 cccacctcgc gggcaaacgt gcgcgccagg tcgcgtatct cggcgatgga cccggcggtt 1260 gtgacgcggg ttgggatcat cccggcggtg aggcgcaaca gggcgtctcg acacccgacg 1320 ggcgactgat cgtaatccag gacaaataga tgcatcggaa ggaggcggtc ggccaagacg 1380 tccaagaccc aggcaaaaat gtggtacaag tccccgttgg gggccagcag ctcgggaacg 1440 cggaacaggg caaacagcgt gtcctcgatg cggggcagag accccgcgcc gtcctcgggg 1500 tcggggcgcg gggtcgccgc ggcgaccccc gtcagccggc cccagtcctc ccgccacctc 1560 ccgccgcgct gcaggtaccg caccgtgttg gcgagtagat cgt 1603 <210> SEQ ID NO 37 <211> LENGTH: 1131 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 37 atggcttctc acgccggcca acagcacgcg cctgcgttcg gtcaggctgc tcgtgcgagc 60 gggcctaccg acggccgcgc ggcgtcccgt cctagccatc gccagggggc ctccggagcc 120 cgcggggatc cggagctgcc cacgctgctg cgggtttata tagacggacc ccacggggtg 180 gggaagacca ccacctccgc gcagctgatg gaggccctgg ggccgcgcga caatatcgtc 240 tacgtccccg agccgatgac ttactggcag gtgctggggg cctccgagac cctgacgaac 300 atctacaaca cgcagcaccg tctggaccgc ggcgagatat cggccgggga ggcggcggtg 360 gtaatgacca gcgcccagat aacaatgagc acgccttatg cggcgacgga cgccgttttg 420 gctcctcata tcggggggga ggctgtgggc ccgcaagccc cgcccccggc cctcaccctt 480 gttttcgacc ggcaccctat cgcctccctg ctgtgctacc cggccgcgcg gtacctcatg 540 ggaagcatga ccccccaggc cgtgttggcg ttcgtggccc tcatgccccc gaccgcgccc 600 ggcacgaacc tggtcctggg tgtccttccg gaggccgaac acgccgaccg cctggccaga 660 cgccaacgcc cgggcgagcg gcttgacctg gccatgctgt ccgccattcg ccgtgtctac 720 gatctactcg ccaacacggt gcggtacctg cagcgcggcg ggaggtggcg ggaggactgg 780 ggccggctga cgggggtcgc cgcggcgacc ccgcgccccg accccgagga cggcgcgggg 840 tctctgcccc gcatcgagga cacgctgttt gccctgttcc gcgttcccga gctgctggcc 900 cccaacgggg acttgtacca catttttgcc tgggtcttgg acgtcttggc cgaccgcctc 960 cttccgatgc atctatttgt cctggattac gatcagtcgc ccgtcgggtg tcgagacgcc 1020 ctgttgcgcc tcaccgccgg gatgatccca acccgcgtca caaccgccgg gtccatcgcc 1080 gagatacgcg acctggcgcg cacgtttgcc cgcgaggtgg ggggagttta g 1131 <210> SEQ ID NO 38 <211> LENGTH: 2517 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 38 atgggccccg gtctgtgggt ggtgatgggg gtcctggtgg gcgttgccgg gggccatgac 60 acgtactgga cggagcaaat cgacccgtgg tttttgcacg gtctggggtt ggcccgcacg 120 tactggcgcg acacaaacac cgggcgtctg tggttgccca acacccccga cgccagcgac 180 ccccagcgcg gacgcttggc gcccccgggc gaactcaacc tgactacggc atccgtgccc 240 atgcttcggt ggtacgccga gcgcttttgt ttcgtgttgg tcaccacggc cgagtttcct 300 cgggaccccg ggcagctgct ttacatccca aagacctatc tgctcggccg gcctcggaac 360 gcgagcctgc ccgagctccc cgaggcgggg cccacgtccc gtccccccgc cgaggtgacc 420 cagctcaagg gactgtcgca caaccccggc gcctccgcgc tgttgcggtc ccgggcctgg 480 gtaacattcg cggccgcgcc ggaccgcgag gggcttacgt tcccgcgggg agacgacggg 540 gcgaccgaga ggcacccgga cggccgacgc aacgcgccgc ccccggggcc gcccgcgggg 600 acaccgaggc atccgacgac gaacctgagc atcgcgcatc tgcacaacgc atccgtgacc 660 tggctggccg ccaggggcct gctacggact ccgggtcggt acgtgtacct ctccccgtcg 720 gcctcgacgt ggcccgtggg cgtctggacg acgggcgggc tggcgttcgg gtgcgacgcc 780 gcgctcgtgc gcgcgcgata cgggaagggc ttcatggggc tcgtgatatc gatgcgggac 840 agccctccgg ccgagatcat agtggtgcct gcggacaaga ccctcgctcg ggtcggaaat 900 ccgaccgacg aaaacgcccc cgcggtgctc cccgggcctc cggccggccc caggtatcgc 960 gtctttgtcc tgggggcccc gacgcccgcc gacaacggct cggcgctgga cgccctccgg 1020 cgggtggccg gctaccccga ggagagcacg aactacgccc agtatatgtc gcgggcctat 1080 gcggagtttt tgggggagga cccgggctcc ggcacggacg cgcgtccgtc cctgttctgg 1140 cgcctcgcgg ggctgctcgc ctcgtcgggg tttgcgttcg tcaacgcggc ccacgcccac 1200 gacgcgattc gcctctccga cctgctgggc tttttggccc actcgcgcgt gctggccggc 1260 ctggccgccc ggggagcagc gggctgcgcg gccgactcgg tgttcctgaa cgtgtccgtg 1320 ttggacccgg cggcccgcct gcggctggag gcgcgcctcg ggcatctggt ggccgcgatc 1380 ctcgagcgag agcagagcct ggtggcgcac gcgctgggct atcagctggc gttcgtgttg 1440 gacagccccg cggcctatgg cgcggtggcc ccgagcgcgg cccgcctgat cgacgccctg 1500 tacgccgagt ttctcggcgg ccgcgcgcta accgccccga tggtccgccg agcgctgttt 1560 tacgccacgg ccgtcctccg ggcgccgttc ctggcgggcg cgccctcggc cgagcagcgg 1620 gaacgcgccc gccggggcct cctcataacc acggccctgt gtacgtccga cgtcgccgcg 1680 gcgacccacg ccgatctccg ggccgcgcta gccaggaccg accaccagaa aaacctcttc 1740 tggctcccgg accacttttc cccatgcgca gcttccctgc gcttcgatct cgccgagggc 1800 gggttcatcc tggacgcgct ggccatggcc acccgatccg acatcccggc ggacgtcatg 1860 gcacaacaga cccgcggcgt ggcctccgtt ctcacgcgct gggcgcacta caacgccctg 1920 atccgcgcct tcgtcccgga ggccacccac cagtgtagcg gcccgtcgca caacgcggag 1980 ccccggatcc tcgtgcccat cacccacaac gccagctacg tcgtcaccca cacccccttg 2040 ccccgcggga tcggatacaa gcttacgggc gttgacgtcc gccgcccgct gtttatcacc 2100 tatctcaccg ccacctgcga agggcacgcg cgggagattg agccgaagcg gctggtgcgc 2160 accgaaaacc ggcgcgacct cggcctcgtg ggggccgtgt ttctgcgcta caccccggcc 2220 ggggaggtca tgtcggtgct gctggtggac acggatgcca cccaacagca gctggcccag 2280 gggccggtgg cgggcacccc gaacgtgttt tccagcgacg tgccgtccgt ggccctgttg 2340 ttgttcccca acggaactgt gattcatctg ctggcctttg acacgctgcc catcgccacc 2400 atcgcccccg ggtttctggc cgcgtccgcg ctgggggtcg ttatgattac cgcggccctg 2460 gcgggcatcc ttagggtggt ccgaacgtgc gtcccatttt tgtggagacg cgaataa 2517 <210> SEQ ID NO 39 <211> LENGTH: 376 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 39 Met Ala Ser His Ala Gly Gln Gln His Ala Pro Ala Phe Gly Gln Ala 5 10 15 Ala Arg Ala Ser Gly Pro Thr Asp Gly Arg Ala Ala Ser Arg Pro Ser 20 25 30 His Arg Gln Gly Ala Ser Gly Ala Arg Gly Asp Pro Glu Leu Pro Thr 35 40 45 Leu Leu Arg Val Tyr Ile Asp Gly Pro His Gly Val Gly Lys Thr Thr 50 55 60 Thr Ser Ala Gln Leu Met Glu Ala Leu Gly Pro Arg Asp Asn Ile Val 65 70 75 80 Tyr Val Pro Glu Pro Met Thr Tyr Trp Gln Val Leu Gly Ala Ser Glu 85 90 95 Thr Leu Thr Asn Ile Tyr Asn Thr Gln His Arg Leu Asp Arg Gly Glu 100 105 110 Ile Ser Ala Gly Glu Ala Ala Val Val Met Thr Ser Ala Gln Ile Thr 115 120 125 Met Ser Thr Pro Tyr Ala Ala Thr Asp Ala Val Leu Ala Pro His Ile 130 135 140 Gly Gly Glu Ala Val Gly Pro Gln Ala Pro Pro Pro Ala Leu Thr Leu 145 150 155 160 Val Phe Asp Arg His Pro Ile Ala Ser Leu Leu Cys Tyr Pro Ala Ala 165 170 175 Arg Tyr Leu Met Gly Ser Met Thr Pro Gln Ala Val Leu Ala Phe Val 180 185 190 Ala Leu Met Pro Pro Thr Ala Pro Gly Thr Asn Leu Val Leu Gly Val 195 200 205 Leu Pro Glu Ala Glu His Ala Asp Arg Leu Ala Arg Arg Gln Arg Pro 210 215 220 Gly Glu Arg Leu Asp Leu Ala Met Leu Ser Ala Ile Arg Arg Val Tyr 225 230 235 240 Asp Leu Leu Ala Asn Thr Val Arg Tyr Leu Gln Arg Gly Gly Arg Trp 245 250 255 Arg Glu Asp Trp Gly Arg Leu Thr Gly Val Ala Ala Ala Thr Pro Arg 260 265 270 Pro Asp Pro Glu Asp Gly Ala Gly Ser Leu Pro Arg Ile Glu Asp Thr 275 280 285 Leu Phe Ala Leu Phe Arg Val Pro Glu Leu Leu Ala Pro Asn Gly Asp 290 295 300 Leu Tyr His Ile Phe Ala Trp Val Leu Asp Val Leu Ala Asp Arg Leu 305 310 315 320 Leu Pro Met His Leu Phe Val Leu Asp Tyr Asp Gln Ser Pro Val Gly 325 330 335 Cys Arg Asp Ala Leu Leu Arg Leu Thr Ala Gly Met Ile Pro Thr Arg 340 345 350 Val Thr Thr Ala Gly Ser Ile Ala Glu Ile Arg Asp Leu Ala Arg Thr 355 360 365 Phe Ala Arg Glu Val Gly Gly Val 370 375 <210> SEQ ID NO 40 <211> LENGTH: 136 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 40 Asp Leu Leu Ala Asn Thr Val Arg Tyr Leu Gln Arg Gly Gly Arg Trp 5 10 15 Arg Glu Asp Trp Gly Arg Leu Thr Gly Val Ala Ala Ala Thr Pro Arg 20 25 30 Pro Asp Pro Glu Asp Gly Ala Gly Ser Leu Pro Arg Ile Glu Asp Thr 35 40 45 Leu Phe Ala Leu Phe Arg Val Pro Glu Leu Leu Ala Pro Asn Gly Asp 50 55 60 Leu Tyr His Ile Phe Ala Trp Val Leu Asp Val Leu Ala Asp Arg Leu 65 70 75 80 Leu Pro Met His Leu Phe Val Leu Asp Tyr Asp Gln Ser Pro Val Gly 85 90 95 Cys Arg Asp Ala Leu Leu Arg Leu Thr Ala Gly Met Ile Pro Thr Arg 100 105 110 Val Thr Thr Ala Gly Ser Ile Ala Glu Ile Arg Asp Leu Ala Arg Thr 115 120 125 Phe Ala Arg Glu Val Gly Gly Val 130 135 <210> SEQ ID NO 41 <211> LENGTH: 284 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 41 Met Gly Pro Gly Leu Trp Val Val Met Gly Val Leu Val Gly Val Ala 5 10 15 Gly Gly His Asp Thr Tyr Trp Thr Glu Gln Ile Asp Pro Trp Phe Leu 20 25 30 His Gly Leu Gly Leu Ala Arg Thr Tyr Trp Arg Asp Thr Asn Thr Gly 35 40 45 Arg Leu Trp Leu Pro Asn Thr Pro Asp Ala Ser Asp Pro Gln Arg Gly 50 55 60 Arg Leu Ala Pro Pro Gly Glu Leu Asn Leu Thr Thr Ala Ser Val Pro 65 70 75 80 Met Leu Arg Trp Tyr Ala Glu Arg Phe Cys Phe Val Leu Val Thr Thr 85 90 95 Ala Glu Phe Pro Arg Asp Pro Gly Gln Leu Leu Tyr Ile Pro Lys Thr 100 105 110 Tyr Leu Leu Gly Arg Pro Arg Asn Ala Ser Leu Pro Glu Leu Pro Glu 115 120 125 Ala Gly Pro Thr Ser Arg Pro Pro Ala Glu Val Thr Gln Leu Lys Gly 130 135 140 Leu Ser His Asn Pro Gly Ala Ser Ala Leu Leu Arg Ser Arg Ala Trp 145 150 155 160 Val Thr Phe Ala Ala Ala Pro Asp Arg Glu Gly Leu Thr Phe Pro Arg 165 170 175 Gly Asp Asp Gly Ala Thr Glu Arg His Pro Asp Gly Arg Arg Asn Ala 180 185 190 Pro Pro Pro Gly Pro Pro Ala Gly Thr Pro Arg His Pro Thr Thr Asn 195 200 205 Leu Ser Ile Ala His Leu His Asn Ala Ser Val Thr Trp Leu Ala Ala 210 215 220 Arg Gly Leu Leu Arg Thr Pro Gly Arg Tyr Val Tyr Leu Ser Pro Ser 225 230 235 240 Ala Ser Thr Trp Pro Val Gly Val Trp Thr Thr Gly Gly Leu Ala Phe 245 250 255 Gly Cys Asp Ala Ala Leu Val Arg Ala Arg Tyr Gly Lys Gly Phe Met 260 265 270 Gly Leu Val Ile Ser Met Arg Asp Ser Pro Pro Ala 275 280 <210> SEQ ID NO 42 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 42 Ser Leu Pro Arg Ile Glu Asp Thr Leu Phe Ala Leu Phe Arg Val 5 10 15 <210> SEQ ID NO 43 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 43 Gly Ser Ile Ala Glu Ile Arg Asp Leu Ala Arg Thr Phe Ala Arg 5 10 15 <210> SEQ ID NO 44 <211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 44 Glu Ile Arg Asp Leu Ala Arg Thr Phe Ala Arg Glu Val Gly Gly Val 5 10 15 <210> SEQ ID NO 45 <211> LENGTH: 838 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 45 Met Gly Pro Gly Leu Trp Val Val Met Gly Val Leu Val Gly Val Ala 5 10 15 Gly Gly His Asp Thr Tyr Trp Thr Glu Gln Ile Asp Pro Trp Phe Leu 20 25 30 His Gly Leu Gly Leu Ala Arg Thr Tyr Trp Arg Asp Thr Asn Thr Gly 35 40 45 Arg Leu Trp Leu Pro Asn Thr Pro Asp Ala Ser Asp Pro Gln Arg Gly 50 55 60 Arg Leu Ala Pro Pro Gly Glu Leu Asn Leu Thr Thr Ala Ser Val Pro 65 70 75 80 Met Leu Arg Trp Tyr Ala Glu Arg Phe Cys Phe Val Leu Val Thr Thr 85 90 95 Ala Glu Phe Pro Arg Asp Pro Gly Gln Leu Leu Tyr Ile Pro Lys Thr 100 105 110 Tyr Leu Leu Gly Arg Pro Arg Asn Ala Ser Leu Pro Glu Leu Pro Glu 115 120 125 Ala Gly Pro Thr Ser Arg Pro Pro Ala Glu Val Thr Gln Leu Lys Gly 130 135 140 Leu Ser His Asn Pro Gly Ala Ser Ala Leu Leu Arg Ser Arg Ala Trp 145 150 155 160 Val Thr Phe Ala Ala Ala Pro Asp Arg Glu Gly Leu Thr Phe Pro Arg 165 170 175 Gly Asp Asp Gly Ala Thr Glu Arg His Pro Asp Gly Arg Arg Asn Ala 180 185 190 Pro Pro Pro Gly Pro Pro Ala Gly Thr Pro Arg His Pro Thr Thr Asn 195 200 205 Leu Ser Ile Ala His Leu His Asn Ala Ser Val Thr Trp Leu Ala Ala 210 215 220 Arg Gly Leu Leu Arg Thr Pro Gly Arg Tyr Val Tyr Leu Ser Pro Ser 225 230 235 240 Ala Ser Thr Trp Pro Val Gly Val Trp Thr Thr Gly Gly Leu Ala Phe 245 250 255 Gly Cys Asp Ala Ala Leu Val Arg Ala Arg Tyr Gly Lys Gly Phe Met 260 265 270 Gly Leu Val Ile Ser Met Arg Asp Ser Pro Pro Ala Glu Ile Ile Val 275 280 285 Val Pro Ala Asp Lys Thr Leu Ala Arg Val Gly Asn Pro Thr Asp Glu 290 295 300 Asn Ala Pro Ala Val Leu Pro Gly Pro Pro Ala Gly Pro Arg Tyr Arg 305 310 315 320 Val Phe Val Leu Gly Ala Pro Thr Pro Ala Asp Asn Gly Ser Ala Leu 325 330 335 Asp Ala Leu Arg Arg Val Ala Gly Tyr Pro Glu Glu Ser Thr Asn Tyr 340 345 350 Ala Gln Tyr Met Ser Arg Ala Tyr Ala Glu Phe Leu Gly Glu Asp Pro 355 360 365 Gly Ser Gly Thr Asp Ala Arg Pro Ser Leu Phe Trp Arg Leu Ala Gly 370 375 380 Leu Leu Ala Ser Ser Gly Phe Ala Phe Val Asn Ala Ala His Ala His 385 390 395 400 Asp Ala Ile Arg Leu Ser Asp Leu Leu Gly Phe Leu Ala His Ser Arg 405 410 415 Val Leu Ala Gly Leu Ala Ala Arg Gly Ala Ala Gly Cys Ala Ala Asp 420 425 430 Ser Val Phe Leu Asn Val Ser Val Leu Asp Pro Ala Ala Arg Leu Arg 435 440 445 Leu Glu Ala Arg Leu Gly His Leu Val Ala Ala Ile Leu Glu Arg Glu 450 455 460 Gln Ser Leu Val Ala His Ala Leu Gly Tyr Gln Leu Ala Phe Val Leu 465 470 475 480 Asp Ser Pro Ala Ala Tyr Gly Ala Val Ala Pro Ser Ala Ala Arg Leu 485 490 495 Ile Asp Ala Leu Tyr Ala Glu Phe Leu Gly Gly Arg Ala Leu Thr Ala 500 505 510 Pro Met Val Arg Arg Ala Leu Phe Tyr Ala Thr Ala Val Leu Arg Ala 515 520 525 Pro Phe Leu Ala Gly Ala Pro Ser Ala Glu Gln Arg Glu Arg Ala Arg 530 535 540 Arg Gly Leu Leu Ile Thr Thr Ala Leu Cys Thr Ser Asp Val Ala Ala 545 550 555 560 Ala Thr His Ala Asp Leu Arg Ala Ala Leu Ala Arg Thr Asp His Gln 565 570 575 Lys Asn Leu Phe Trp Leu Pro Asp His Phe Ser Pro Cys Ala Ala Ser 580 585 590 Leu Arg Phe Asp Leu Ala Glu Gly Gly Phe Ile Leu Asp Ala Leu Ala 595 600 605 Met Ala Thr Arg Ser Asp Ile Pro Ala Asp Val Met Ala Gln Gln Thr 610 615 620 Arg Gly Val Ala Ser Val Leu Thr Arg Trp Ala His Tyr Asn Ala Leu 625 630 635 640 Ile Arg Ala Phe Val Pro Glu Ala Thr His Gln Cys Ser Gly Pro Ser 645 650 655 His Asn Ala Glu Pro Arg Ile Leu Val Pro Ile Thr His Asn Ala Ser 660 665 670 Tyr Val Val Thr His Thr Pro Leu Pro Arg Gly Ile Gly Tyr Lys Leu 675 680 685 Thr Gly Val Asp Val Arg Arg Pro Leu Phe Ile Thr Tyr Leu Thr Ala 690 695 700 Thr Cys Glu Gly His Ala Arg Glu Ile Glu Pro Lys Arg Leu Val Arg 705 710 715 720 Thr Glu Asn Arg Arg Asp Leu Gly Leu Val Gly Ala Val Phe Leu Arg 725 730 735 Tyr Thr Pro Ala Gly Glu Val Met Ser Val Leu Leu Val Asp Thr Asp 740 745 750 Ala Thr Gln Gln Gln Leu Ala Gln Gly Pro Val Ala Gly Thr Pro Asn 755 760 765 Val Phe Ser Ser Asp Val Pro Ser Val Ala Leu Leu Leu Phe Pro Asn 770 775 780 Gly Thr Val Ile His Leu Leu Ala Phe Asp Thr Leu Pro Ile Ala Thr 785 790 795 800 Ile Ala Pro Gly Phe Leu Ala Ala Ser Ala Leu Gly Val Val Met Ile 805 810 815 Thr Ala Ala Leu Ala Gly Ile Leu Arg Val Val Arg Thr Cys Val Pro 820 825 830 Phe Leu Trp Arg Arg Glu 835 <210> SEQ ID NO 46 <211> LENGTH: 215 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 46 Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser 5 10 15 Ser Ala Gly Gly Ala Gly Gly Ser Val Ala Ser Ala Ser Gly Ala Gly 20 25 30 Glu Arg Arg Glu Thr Ser Leu Gly Pro Arg Ala Ala Ala Pro Arg Gly 35 40 45 Pro Arg Lys Cys Ala Arg Lys Thr Arg His Ala Glu Gly Gly Pro Glu 50 55 60 Pro Gly Ala Arg Asp Pro Ala Pro Gly Leu Thr Arg Tyr Leu Pro Ile 65 70 75 80 Ala Gly Val Ser Ser Val Val Ala Leu Ala Pro Tyr Val Asn Lys Thr 85 90 95 Val Thr Gly Asp Cys Leu Pro Val Leu Asp Met Glu Thr Gly His Ile 100 105 110 Gly Ala Tyr Val Val Leu Val Asp Gln Thr Gly Asn Val Ala Asp Leu 115 120 125 Leu Arg Ala Ala Ala Pro Ala Trp Ser Arg Arg Thr Leu Leu Pro Glu 130 135 140 His Ala Arg Asn Cys Val Arg Pro Pro Asp Tyr Pro Thr Pro Pro Ala 145 150 155 160 Ser Glu Trp Asn Ser Leu Trp Met Thr Pro Val Gly Asn Met Leu Phe 165 170 175 Asp Gln Gly Thr Leu Val Gly Ala Leu Asp Phe His Gly Leu Arg Ser 180 185 190 Arg His Pro Trp Ser Arg Glu Gln Gly Ala Pro Ala Pro Ala Gly Asp 195 200 205 Ala Pro Ala Gly His Gly Glu 210 215 <210> SEQ ID NO 47 <211> LENGTH: 826 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 47 Met Glu Pro Arg Pro Gly Thr Ser Ser Arg Ala Asp Pro Gly Pro Glu 5 10 15 Arg Pro Pro Arg Gln Thr Pro Gly Thr Gln Pro Ala Ala Pro His Ala 20 25 30 Trp Gly Met Leu Asn Asp Met Gln Trp Leu Ala Ser Ser Asp Ser Glu 35 40 45 Glu Glu Thr Glu Val Gly Ile Ser Asp Asp Asp Leu His Arg Asp Ser 50 55 60 Thr Ser Glu Ala Gly Ser Thr Asp Thr Glu Met Phe Glu Ala Gly Leu 65 70 75 80 Met Asp Ala Ala Thr Pro Pro Ala Arg Pro Pro Ala Glu Arg Gln Gly 85 90 95 Ser Pro Thr Pro Ala Asp Ala Gln Gly Ser Cys Gly Gly Gly Pro Val 100 105 110 Gly Glu Glu Glu Ala Glu Ala Gly Gly Gly Gly Asp Val Cys Ala Val 115 120 125 Cys Thr Asp Glu Ile Ala Pro Pro Leu Arg Cys Gln Ser Phe Pro Cys 130 135 140 Leu His Pro Phe Cys Ile Pro Cys Met Lys Thr Trp Ile Pro Leu Arg 145 150 155 160 Asn Thr Cys Pro Leu Cys Asn Thr Pro Val Ala Tyr Leu Ile Val Gly 165 170 175 Val Thr Ala Ser Gly Ser Phe Ser Thr Ile Pro Ile Val Asn Asp Pro 180 185 190 Arg Thr Arg Val Glu Ala Glu Ala Ala Val Arg Ala Gly Thr Ala Val 195 200 205 Asp Phe Ile Trp Thr Gly Asn Pro Arg Thr Ala Pro Arg Ser Leu Ser 210 215 220 Leu Gly Gly His Thr Val Arg Ala Leu Ser Pro Thr Pro Pro Trp Pro 225 230 235 240 Gly Thr Asp Asp Glu Asp Asp Asp Leu Ala Asp Gly Val Asp Tyr Val 245 250 255 Pro Pro Ala Pro Arg Arg Ala Pro Arg Arg Gly Gly Gly Gly Ala Gly 260 265 270 Ala Thr Arg Gly Thr Ser Gln Pro Ala Ala Thr Arg Pro Ala Pro Pro 275 280 285 Gly Ala Pro Arg Ser Ser Ser Ser Gly Gly Ala Pro Leu Arg Ala Gly 290 295 300 Val Gly Ser Gly Ser Gly Gly Gly Pro Ala Val Ala Ala Val Val Pro 305 310 315 320 Arg Val Ala Ser Leu Pro Pro Ala Ala Gly Gly Gly Arg Ala Gln Ala 325 330 335 Arg Arg Val Gly Glu Asp Ala Ala Ala Ala Glu Gly Arg Thr Pro Pro 340 345 350 Ala Arg Gln Pro Arg Ala Ala Gln Glu Pro Pro Ile Val Ile Ser Asp 355 360 365 Ser Pro Pro Pro Ser Pro Arg Arg Pro Ala Gly Pro Gly Pro Leu Ser 370 375 380 Phe Val Ser Ser Ser Ser Ala Gln Val Ser Ser Gly Pro Gly Gly Gly 385 390 395 400 Gly Leu Pro Gln Ser Ser Gly Arg Ala Ala Arg Pro Arg Ala Ala Val 405 410 415 Ala Pro Arg Val Arg Ser Pro Pro Arg Ala Ala Ala Ala Pro Val Val 420 425 430 Ser Ala Ser Ala Asp Ala Ala Gly Pro Ala Pro Pro Ala Val Pro Val 435 440 445 Asp Ala His Arg Ala Pro Arg Ser Arg Met Thr Gln Ala Gln Thr Asp 450 455 460 Thr Gln Ala Gln Ser Leu Gly Arg Ala Gly Ala Thr Asp Ala Arg Gly 465 470 475 480 Ser Gly Gly Pro Gly Ala Glu Gly Gly Pro Gly Val Pro Arg Gly Thr 485 490 495 Asn Thr Pro Gly Ala Ala Pro His Ala Ala Glu Gly Ala Ala Ala Arg 500 505 510 Pro Arg Lys Arg Arg Gly Ser Asp Ser Gly Pro Ala Ala Ser Ser Ser 515 520 525 Ala Ser Ser Ser Ala Ala Pro Arg Ser Pro Leu Ala Pro Gln Gly Val 530 535 540 Gly Ala Lys Arg Ala Ala Pro Arg Arg Ala Pro Asp Ser Asp Ser Gly 545 550 555 560 Asp Arg Gly His Gly Pro Leu Ala Pro Ala Ser Ala Gly Ala Ala Pro 565 570 575 Pro Ser Ala Ser Pro Ser Ser Gln Ala Ala Val Ala Ala Ala Ser Ser 580 585 590 Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser 595 600 605 Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser 610 615 620 Ala Ser Ser Ser Ala Gly Gly Ala Gly Gly Ser Val Ala Ser Ala Ser 625 630 635 640 Gly Ala Gly Glu Arg Arg Glu Thr Ser Leu Gly Pro Arg Ala Ala Ala 645 650 655 Pro Arg Gly Pro Arg Lys Cys Ala Arg Lys Thr Arg His Ala Glu Gly 660 665 670 Gly Pro Glu Pro Gly Ala Arg Asp Pro Ala Pro Gly Leu Thr Arg Tyr 675 680 685 Leu Pro Ile Ala Gly Val Ser Ser Val Val Ala Leu Ala Pro Tyr Val 690 695 700 Asn Lys Thr Val Thr Gly Asp Cys Leu Pro Val Leu Asp Met Glu Thr 705 710 715 720 Gly His Ile Gly Ala Tyr Val Val Leu Val Asp Gln Thr Gly Asn Val 725 730 735 Ala Asp Leu Leu Arg Ala Ala Ala Pro Ala Trp Ser Arg Arg Thr Leu 740 745 750 Leu Pro Glu His Ala Arg Asn Cys Val Arg Pro Pro Asp Tyr Pro Thr 755 760 765 Pro Pro Ala Ser Glu Trp Asn Ser Leu Trp Met Thr Pro Val Gly Asn 770 775 780 Met Leu Phe Asp Gln Gly Thr Leu Val Gly Ala Leu Asp Phe His Gly 785 790 795 800 Leu Arg Ser Arg His Pro Trp Ser Arg Glu Gln Gly Ala Pro Ala Pro 805 810 815 Ala Gly Asp Ala Pro Ala Gly His Gly Glu 820 825 <210> SEQ ID NO 48 <211> LENGTH: 3350 <212> TYPE: DNA <213> ORGANISM: HSV-2 <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 1027, 1034, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061, 1062, 1063, 1064, 1065, 1066, 1067, 1068, 1069, 1070, 1071, 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081, 1082, 1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090 <223> OTHER INFORMATION: n = A,T,C or G <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 1091, 1092, 1093, 1094, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 1106, 1107, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1129 <223> OTHER INFORMATION: n = A,T,C or G <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 1130, 1131, 1132, 1133, 1134, 1135, 1136, 1137, 1138, 1139, 1140, 1141, 1142, 1143, 1144, 1145, 1146, 1147, 1148, 1149, 1150, 1151, 1152, 1327, 1364, 1390, 1392 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 48 ccgtcggtga cctgcaggag ctcgtttatt aatagccagt ccatgctcag cgtagcggcc 60 agcccctggg gagacaggtc cacggagtcc ggaaccaccg tcggctgacc caggggcccc 120 aggctgtagt ccccccaggc ccccaggtca tgacggttcg tgagcacgac gaggtctgcg 180 gccgggctgg ggggcgcgtc ctcggtcgcg tgggccatca cctcctgaat ggctgcggtg 240 cgctgatcgg ccgagctggc gaagggcgcc acgaccagcg cgcgctccgt ctgcaggccc 300 ttccacgtgt cgtggagttc ctgaacgaac tcggccaccc gctcggggcc cgtggccgcg 360 cgcgcggcct gatagccggc cgagaggcgc cgccagcgcg ccaggaactg actcatgtaa 420 cagaacccgg ggacctggtc ccccgacatc aactttgacg ccctggcgtg gatgcccgac 480 acgatggcca ggaacccgtg gatttcccgc cgcacgacgg ccagcacgtt accctcgtgc 540 gagacctggg ccgccagctc gtcgcatacc ccgaggtgcg ccgtcgtctc ggtgacgacg 600 gaccgcagcc ccgcgaggga cgcgaccagc gcgcgcttgg cgtcgtgata catgccgcag 660 tactggctca ccgcgtcgcc catggcctcg gggcgccagg gccccaggcg ctcgtgggcg 720 tctgcgacca cggcgtacag gcggtgcccg tcgctctcga accggcactc aaagaaggcg 780 gcgagcgtgc gcatgtgaag ccgcagcagc acgatcgcgt cctccagctg gcggaccagg 840 gggtcggcgc gctcggcgag ctcctgcagc accccccggg ccgccagggc gtacatgctg 900 atcagcagca ggctgctgcc cacctcggga ggctgggggg gaggcagctg gaccgcgggc 960 cgcagctgct cgacggcccc cctggcgatc acgtacagct cgcgcagcag ctgctcgatg 1020 ttgtcgngcc atcntgcatc gtgggcccga cgcnnnnnnn nnnnnnnnnn nnnnnnnnnn 1080 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1140 nnnnnnnnnn nnaggccagc acccgcaggg caaactcgat ggggcggggc aggtaggcag 1200 cgttgcacgt ggccctcagc gcgtccccga ccaccagggc cagcacgtaa gggacgaacc 1260 ccgggtcggc gaggacgttg gggtggatgc cctccagggc cgggaagcgg atcttggtgg 1320 ccgcggncag gtgaaccgag ggggcgtggc taggcggccc gacngggagc atcgcggaca 1380 gcggcgtggn cngggtggtg ggggtcaggt cccagtgggt ctggccgtac acgtcgagcc 1440 agatgagcgc cgtctcgcgc aggaggctgg gctggccggc gctgaagcgg cgctcggccg 1500 tctcaaactc ccccacgagc gtgcgccgca ggctcgccag gtgttccgtc ggcacggccg 1560 ggcccatgat gcgcgccagc gtctggctga ggacgccgcc cgacaggccg accgcctcac 1620 agagccgccc gtgcgtgtgc tcgctggcgc cctggatccg ccggaacgtt ttcacgtagc 1680 cggcgtagtg cccgtactcc cgcgcgagcc cgaacacgtt cgcccccgca agggcaatgc 1740 acccaaagag ctgctggatc tcgctgagcc cgtggccggg gggcgtccgc gcgggcaccc 1800 ccgccaccaa aaacccctcc agggccgata tgtactgggt gcagtgcgcg ggcgtgaacc 1860 ccgcgtcggt aagcgtgttg atcaccacgg agggcgagtt gctgttctgg accaaagccc 1920 acgtctgctg cagcagcgcg aggagccgtt gctgggcccc ggcggagggc ggctccccta 1980 gctgcagcag gccggtgacg gccggacgga agatggccag cgccgacgca ctcagaaacg 2040 gcacgtcggg gtcgaagacg gccgcgtccg tccgcacgcg cgccatcagc gtccccgggg 2100 gcgcgcacgc cgaccgcggg ctgacgcggc ttagggcggt cgacacgcgc acctcctcgc 2160 gactgcgaac cattttggtg gcctcgaggg gcgggatcat gatagccggg tcgatctccc 2220 gcaccgtgtg ctgaaactgg gccagcagcg gcggcgggac caccgcgccc cgatcggggg 2280 tcgtcaggta ctcgtccacc agcgccagcg taaacagggc ccgcgtgagg ggggtcaggg 2340 cggcgtcgtc gatgcgctgt aggtgcgccg agaacagcgt cacccaattg ctgaccaggg 2400 ccaagaaccg gagaccctct tgcacgatcg gggacgggaa gagcaggctg tacgccgggg 2460 tggtcaggtt ggcgccgggt tgccccaggg gaaccgggga catcttaagc gacatctccc 2520 cgagggcctc cagggaggtc cgcgggttca tggccaggca gctctgggtg acggtccgcc 2580 agcggtcgat ccactccacg gcacactggc ggacgcgcac cggccccagg gccgccgtgg 2640 tgcgcagccc ggcggcctcc agcgcgtggg tcgtgtcgga gccggtgatc gccaggaccg 2700 tgtccttgat gacgtccatc tcccggaagg ccgcctcggg ggtctcgggg agcgccaccg 2760 ccatgcggtg caccagcagc ccggggaggt tctcggccaa gagcgccgtc tccggaagcc 2820 cgtgggcccg gtgcaaggcg cacagttgct ccaggagcgg gtgccagcac gcccgcgcct 2880 ccgccgggcc gaccgccgcg cccgacaaca gaaacgccgc cgtggcggcg cgcagtttgg 2940 ccgcggacag aaacgccggc tcgtccgcgc tgcccgccgg ctcgctcgag ggggagggcg 3000 gccggcggag gttggtcagg ctccccaaca ggacctgcaa cggtccgttt gggggtggag 3060 cggacggggg ggtcatgccg gcgggcgccg ggacctggag cgcgctgtcc gacatggcga 3120 ccggcgtgcg cgctcggcga cgcggcgcgg agaccgcggg cccaaacggg aatgactgcc 3180 gccgccctat acggaggggc taagtatcgc ccggggaccc ttcgaaaccc cgggcgtgtc 3240 gcaagtacgc cgcgaaggcg cggcgtgtta tacggcgcgt tatgtcccgg cattccgttc 3300 gtgggttcgg gcccgggtgc tgtcgggtgg gagtgtgtgt gtgtgggggg 3350 <210> SEQ ID NO 49 <211> LENGTH: 3345 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 49 atgtcggaca gcgcgctcca ggtcccggcg cccgccggca tgaccccccc gtccgctcca 60 cccccaaacg gaccgttgca ggtcctgttg gggagcctga ccaacctccg ccggccgccc 120 tccccctcga gcgagccggc gggcagcgcg gacgagccgg cgtttctgtc cgcggccaaa 180 ctgcacgccg ccacggcggc gtttctgttg tcgggcgcgg cggtcggccc ggcggaggcg 240 cgggcgtgct ggcacccgct cctggagcaa ctgtgcgcct tgcaccgggc ccacgggctt 300 ccggagacgg cgctcttggc cgagaacctc cccgggctgc tggtgcaccg catggcggtg 360 gcgctccccg agacccccga ggcggccttc cgggagatgg acgtcatcaa ggacacggtc 420 ctggcgatca ccggctccga cacgacccac gcgctggagg ccgccgggct gcgcaccacg 480 gcggccctgg ggccggtgcg cgtccgccag tgtgccgtgg agtggatcga ccgctggcgg 540 accgtcaccc agagctgcct ggccatgaac ccgcggacct ccctggaggc cctcggggag 600 atgtcgctta agatgtcccc ggttcccctg gggcaacccg gcgccaacct gaccaccccg 660 gcgtacagcc tgctcttccc gtccccgatc gtgcaagagg gtctccggtt cttggccctg 720 gtcagcaatt gggtgacgct gttctcggcg cacctacagc gcatcgacga cgccgccctg 780 acccccctca cgcgggccct gtttacgctg gcgctggtgg acgactacct gacgaccccc 840 gatcggggcg cggtggtccc gccgccgctg ctggcccagt ttcagcacac ggtgcgggag 900 atcgacccgg ctatcatgat cccgcccctc gaggccacca aaatggttcg cagtcgcgag 960 gaggtgcgcg tgtcgaccgc cctaagccgc gtcagcccgc ggtcggcgtg cgcgcccccg 1020 gggacgctga tggcgcgcgt gcggacggac gcggccgtct tcgaccccga cgtgccgttt 1080 ctgagtgcgt cggcgctggc catcttccgt ccggccgtca ccggcctgct gcagctaggg 1140 gagccgccct ccgccggggc ccagcaacgg ctcctcgcgc tgctgcagca gacgtgggct 1200 ttggtccaga acagcaactc gccctccgtg gtgatcaaca cgcttaccga cgcggggttc 1260 acgcccgcgc actgcaccca gtacatatcg gccctggagg ggtttttggt ggcgggggtg 1320 cccgcgcgga cgccccccgg ccacgggctc agcgagatcc agcagctctt tgggtgcatt 1380 gcccttgcgg gggcgaacgt gttcgggctc gcgcgggagt acgggcacta cgccggctac 1440 gtgaaaacgt tccggcggat ccagggcgcc agcgagcaca cgcacgggcg gctctgtgag 1500 gcggtcggcc tgtcgggcgg cgtcctcagc cagacgctgg cgcgcatcat gggcccggcc 1560 gtgccgacgg aacacctggc gagcctgcgg cgcacgctcg tgggggagtt tgagacggcc 1620 gagcgccgct tcagcgccgg ccagcccagc ctcctgcgcg agacggcgct catctggctc 1680 gacgtgtacg gccagaccca ctgggacctg acccccacca ccccggccac gccgctgtcc 1740 gcgctgctcc ccgtcgggcc gcctagccac gccccctcgg ttcacctggc cgcggccacc 1800 aagatccgct tcccggccct ggagggcatc caccccaacg tcctcgccga cccggggttc 1860 gtcccttacg tgctggccct ggtggtcggg gacgcgctga gggccacgtg caacgctgcc 1920 tacctgcccc gccccatcga gtttgccctg cgggtgctgg cctgggcgcg cgacttcggc 1980 ctgggctatc tccccaccgt cgaggggcac cgcacaaaat tgggcgcgct gatcaccctc 2040 ctcgaaccgg ccacccgggc cggcgtcggg cccacgatgc agatggccga caacatcgag 2100 cagctgctgc gcgagctgta cgtgatcgcc aggggggccg tcgagcagct gcggcccgcg 2160 gtccagctgc ctccccccca gcctcccgag gtgggcagca gcctgctgct gatcagcatg 2220 tacgccctgg cggcccgggg ggtgctgcag gagctcgccg agcgcgccga ccccctggtc 2280 cgccagctgg aggacgcgat cgtgctgctg cggctgcaca tgcgcacgct cgccgccttc 2340 tttgagtgcc ggttcgagag cgacgggcac cgcctgtacg ccgtggtcgc agacgcccac 2400 gagcgcctgg ggccctggcg ccccgaggcc atgggcgacg cggtgagcca gtactgcggc 2460 atgtatcacg acgccaagcg cgcgctggtc gcgtccctcg cggggctgcg gtccgtcgtc 2520 accgagacga cggcgcacct cggggtatgc gacgagctgg cggcccaggt ctcgcacgag 2580 ggtaacgtgc tggccgtcgt gcggcgggaa atccacgggt tcctggccat cgtgtcgggc 2640 atccacgcca gggcgtcaaa gttgatgtcg ggggaccagg tccccgggtt ctgttacatg 2700 agtcagttcc tggcgcgctg gcggcgcctc tcggccggct atcaggccgc acgcgcggcc 2760 acgggccccg agcgggtggc cgagttcgtt caggaactcc acgacacgtg gaagggcctg 2820 cagacggagc gcgcgctggt cgtggcgcgc ttcgccagct cggccgatca gcgcaccgca 2880 gccattcagg aggtgatggc ccacgcgacc gaggacgcgc cccccagccc ggccgcagac 2940 ctcgtcgtgc tcacgaaccg tcatgacctg ggggcctggg gggactacag cctggggccc 3000 ctgggtcagc cgacggtggt tccggactcc gtggacctgt ctccccaggg gctggccgct 3060 acgctgagca tggactggct attaataaac gagctcctgc aggtcaccga cggcgtgttt 3120 cgcgcctcgg cgtttcggcc ttccgccggc ccgggggccc ccggggacct ggaggcccaa 3180 gatgccggcg gtagcacccc cgaacccacg acacccggcc cacaggacac gcaggcccgc 3240 gcgccgtcga cgcgcccggc gggccgcgag acggtccctt ggcccaacac ccccgtggag 3300 gacgacgaga tgacgccgca ggagacacca ccggtacacc cgtag 3345 <210> SEQ ID NO 50 <211> LENGTH: 993 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 50 Glu Pro Ala Gly Ser Ala Asp Glu Pro Ala Phe Leu Ser Ala Ala Lys 5 10 15 Leu His Ala Ala Thr Ala Ala Phe Leu Leu Ser Gly Ala Ala Val Gly 20 25 30 Pro Ala Glu Ala Arg Ala Cys Trp His Pro Leu Leu Glu Gln Leu Cys 35 40 45 Ala Leu His Arg Ala His Gly Leu Pro Glu Thr Ala Leu Leu Ala Glu 50 55 60 Asn Leu Pro Gly Leu Leu Val His Arg Met Ala Val Ala Leu Pro Glu 65 70 75 80 Thr Pro Glu Ala Ala Phe Arg Glu Met Asp Val Ile Lys Asp Thr Val 85 90 95 Leu Ala Ile Thr Gly Ser Asp Thr Thr His Ala Leu Glu Ala Ala Gly 100 105 110 Leu Arg Thr Thr Ala Ala Leu Gly Pro Val Arg Val Arg Gln Cys Ala 115 120 125 Val Glu Trp Ile Asp Arg Trp Arg Thr Val Thr Gln Ser Cys Leu Ala 130 135 140 Met Asn Pro Arg Thr Ser Leu Glu Ala Leu Gly Glu Met Ser Leu Lys 145 150 155 160 Met Ser Pro Val Pro Leu Gly Gln Pro Gly Ala Asn Leu Thr Thr Pro 165 170 175 Ala Tyr Ser Leu Leu Phe Pro Ser Pro Ile Val Gln Glu Gly Leu Arg 180 185 190 Phe Leu Ala Leu Val Ser Asn Trp Val Thr Leu Phe Ser Ala His Leu 195 200 205 Gln Arg Ile Asp Asp Ala Ala Leu Thr Pro Leu Thr Arg Ala Leu Phe 210 215 220 Thr Leu Ala Leu Val Asp Asp Tyr Leu Thr Thr Pro Asp Arg Gly Ala 225 230 235 240 Val Val Pro Pro Pro Leu Leu Ala Gln Phe Gln His Thr Val Arg Glu 245 250 255 Ile Asp Pro Ala Ile Met Ile Pro Pro Leu Glu Ala Thr Lys Met Val 260 265 270 Arg Ser Arg Glu Glu Val Arg Val Ser Thr Ala Leu Ser Arg Val Ser 275 280 285 Pro Arg Ser Ala Cys Ala Pro Pro Gly Thr Leu Met Ala Arg Val Arg 290 295 300 Thr Asp Ala Ala Val Phe Asp Pro Asp Val Pro Phe Leu Ser Ala Ser 305 310 315 320 Ala Leu Ala Ile Phe Arg Pro Ala Val Thr Gly Leu Leu Gln Leu Gly 325 330 335 Glu Pro Pro Ser Ala Gly Ala Gln Gln Arg Leu Leu Ala Leu Leu Gln 340 345 350 Gln Thr Trp Ala Leu Val Gln Asn Ser Asn Ser Pro Ser Val Val Ile 355 360 365 Asn Thr Leu Thr Asp Ala Gly Phe Thr Pro Ala His Cys Thr Gln Tyr 370 375 380 Ile Ser Ala Leu Glu Gly Phe Leu Val Ala Gly Val Pro Ala Arg Thr 385 390 395 400 Pro Pro Gly His Gly Leu Ser Glu Ile Gln Gln Leu Phe Gly Cys Ile 405 410 415 Ala Leu Ala Gly Ala Asn Val Phe Gly Leu Ala Arg Glu Tyr Gly His 420 425 430 Tyr Ala Gly Tyr Val Lys Thr Phe Arg Arg Ile Gln Gly Ala Ser Glu 435 440 445 His Thr His Gly Arg Leu Cys Glu Ala Val Gly Leu Ser Gly Gly Val 450 455 460 Leu Ser Gln Thr Leu Ala Arg Ile Met Gly Pro Ala Val Pro Thr Glu 465 470 475 480 His Leu Ala Ser Leu Arg Arg Thr Leu Val Gly Glu Phe Glu Thr Ala 485 490 495 Glu Arg Arg Phe Ser Ala Gly Gln Pro Ser Leu Leu Arg Glu Thr Ala 500 505 510 Leu Ile Trp Leu Asp Val Tyr Gly Gln Thr His Trp Asp Leu Thr Pro 515 520 525 Thr Thr Pro Ala Thr Pro Leu Ser Ala Leu Leu Pro Val Gly Pro Pro 530 535 540 Ser His Ala Pro Ser Val His Leu Ala Ala Ala Thr Lys Ile Arg Phe 545 550 555 560 Pro Ala Leu Glu Gly Ile His Pro Asn Val Leu Ala Asp Pro Gly Phe 565 570 575 Val Pro Tyr Val Leu Ala Leu Val Val Gly Asp Ala Leu Arg Ala Thr 580 585 590 Cys Asn Ala Ala Tyr Leu Pro Arg Pro Ile Glu Phe Ala Leu Arg Val 595 600 605 Leu Ala Trp Ala Arg Asp Phe Gly Leu Gly Tyr Leu Pro Thr Val Glu 610 615 620 Gly His Arg Thr Lys Leu Gly Ala Leu Ile Thr Leu Leu Glu Pro Ala 625 630 635 640 Thr Arg Ala Gly Val Gly Pro Thr Met Gln Met Ala Asp Asn Ile Glu 645 650 655 Gln Leu Leu Arg Glu Leu Tyr Val Ile Ala Arg Gly Ala Val Glu Gln 660 665 670 Leu Arg Pro Ala Val Gln Leu Pro Pro Pro Gln Pro Pro Glu Val Gly 675 680 685 Ser Ser Leu Leu Leu Ile Ser Met Tyr Ala Leu Ala Ala Arg Gly Val 690 695 700 Leu Gln Glu Leu Ala Glu Arg Ala Asp Pro Leu Val Arg Gln Leu Glu 705 710 715 720 Asp Ala Ile Val Leu Leu Arg Leu His Met Arg Thr Leu Ala Ala Phe 725 730 735 Phe Glu Cys Arg Phe Glu Ser Asp Gly His Arg Leu Tyr Ala Val Val 740 745 750 Ala Asp Ala His Glu Arg Leu Gly Pro Trp Arg Pro Glu Ala Met Gly 755 760 765 Asp Ala Val Ser Gln Tyr Cys Gly Met Tyr His Asp Ala Lys Arg Ala 770 775 780 Leu Val Ala Ser Leu Ala Gly Leu Arg Ser Val Val Thr Glu Thr Thr 785 790 795 800 Ala His Leu Gly Val Cys Asp Glu Leu Ala Ala Gln Val Ser His Glu 805 810 815 Gly Asn Val Leu Ala Val Val Arg Arg Glu Ile His Gly Phe Leu Ala 820 825 830 Ile Val Ser Gly Ile His Ala Arg Ala Ser Lys Leu Met Ser Gly Asp 835 840 845 Gln Val Pro Gly Phe Cys Tyr Met Ser Gln Phe Leu Ala Arg Trp Arg 850 855 860 Arg Leu Ser Ala Gly Tyr Gln Ala Ala Arg Ala Ala Thr Gly Pro Glu 865 870 875 880 Arg Val Ala Glu Phe Val Gln Glu Leu His Asp Thr Trp Lys Gly Leu 885 890 895 Gln Thr Glu Arg Ala Leu Val Val Ala Arg Phe Ala Ser Ser Ala Asp 900 905 910 Gln Arg Thr Ala Ala Ile Gln Glu Val Met Ala His Ala Thr Glu Asp 915 920 925 Ala Pro Pro Ser Pro Ala Ala Asp Leu Val Val Leu Thr Asn Arg His 930 935 940 Asp Leu Gly Ala Trp Gly Asp Tyr Ser Leu Gly Pro Leu Gly Gln Pro 945 950 955 960 Thr Val Val Pro Asp Ser Val Asp Leu Ser Pro Gln Gly Leu Ala Ala 965 970 975 Thr Leu Ser Met Asp Trp Leu Leu Ile Asn Glu Leu Leu Gln Val Thr 980 985 990 Asp <210> SEQ ID NO 51 <211> LENGTH: 1113 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 51 Met Ser Asp Ser Ala Leu Gln Val Pro Ala Pro Ala Gly Met Thr Pro 5 10 15 Pro Ser Ala Pro Pro Pro Asn Gly Pro Leu Gln Val Leu Leu Gly Ser 20 25 30 Leu Thr Asn Leu Arg Arg Pro Pro Ser Pro Ser Ser Glu Pro Ala Gly 35 40 45 Ser Ala Asp Glu Pro Ala Phe Leu Ser Ala Ala Lys Leu His Ala Ala 50 55 60 Thr Ala Ala Phe Leu Leu Ser Gly Ala Ala Val Gly Pro Ala Glu Ala 65 70 75 80 Arg Ala Cys Trp His Pro Leu Leu Glu Gln Leu Cys Ala Leu His Arg 85 90 95 Ala His Gly Leu Pro Glu Thr Ala Leu Leu Ala Glu Asn Leu Pro Gly 100 105 110 Leu Leu Val His Arg Met Ala Val Ala Leu Pro Glu Thr Pro Glu Ala 115 120 125 Ala Phe Arg Glu Met Asp Val Ile Lys Asp Thr Val Leu Ala Ile Thr 130 135 140 Gly Ser Asp Thr Thr His Ala Leu Glu Ala Ala Gly Leu Arg Thr Thr 145 150 155 160 Ala Ala Leu Gly Pro Val Arg Val Arg Gln Cys Ala Val Glu Trp Ile 165 170 175 Asp Arg Trp Arg Thr Val Thr Gln Ser Cys Leu Ala Met Asn Pro Arg 180 185 190 Thr Ser Leu Glu Ala Leu Gly Glu Met Ser Leu Lys Met Ser Pro Val 195 200 205 Pro Leu Gly Gln Pro Gly Ala Asn Leu Thr Thr Pro Ala Tyr Ser Leu 210 215 220 Leu Phe Pro Ser Pro Ile Val Gln Glu Gly Leu Arg Phe Leu Ala Leu 225 230 235 240 Val Ser Asn Trp Val Thr Leu Phe Ser Ala His Leu Gln Arg Ile Asp 245 250 255 Asp Ala Ala Leu Thr Pro Leu Thr Arg Ala Leu Phe Thr Leu Ala Leu 260 265 270 Val Asp Asp Tyr Leu Thr Thr Pro Asp Arg Gly Ala Val Val Pro Pro 275 280 285 Pro Leu Leu Ala Gln Phe Gln His Thr Val Arg Glu Ile Asp Pro Ala 290 295 300 Ile Met Ile Pro Pro Leu Glu Ala Thr Lys Met Val Arg Ser Arg Glu 305 310 315 320 Glu Val Arg Val Ser Thr Ala Leu Ser Arg Val Ser Pro Arg Ser Ala 325 330 335 Cys Ala Pro Pro Gly Thr Leu Met Ala Arg Val Arg Thr Asp Ala Ala 340 345 350 Val Phe Asp Pro Asp Val Pro Phe Leu Ser Ala Ser Ala Leu Ala Ile 355 360 365 Phe Arg Pro Ala Val Thr Gly Leu Leu Gln Leu Gly Glu Pro Pro Ser 370 375 380 Ala Gly Ala Gln Gln Arg Leu Leu Ala Leu Leu Gln Gln Thr Trp Ala 385 390 395 400 Leu Val Gln Asn Ser Asn Ser Pro Ser Val Val Ile Asn Thr Leu Thr 405 410 415 Asp Ala Gly Phe Thr Pro Ala His Cys Thr Gln Tyr Ile Ser Ala Leu 420 425 430 Glu Gly Phe Leu Val Ala Gly Val Pro Ala Arg Thr Pro Pro Gly His 435 440 445 Gly Leu Ser Glu Ile Gln Gln Leu Phe Gly Cys Ile Ala Leu Ala Gly 450 455 460 Ala Asn Val Phe Gly Leu Ala Arg Glu Tyr Gly His Tyr Ala Gly Tyr 465 470 475 480 Val Lys Thr Phe Arg Arg Ile Gln Gly Ala Ser Glu His Thr His Gly 485 490 495 Arg Leu Cys Glu Ala Val Gly Leu Ser Gly Gly Val Leu Ser Gln Thr 500 505 510 Leu Ala Arg Ile Met Gly Pro Ala Val Pro Thr Glu His Leu Ala Ser 515 520 525 Leu Arg Arg Thr Leu Val Gly Glu Phe Glu Thr Ala Glu Arg Arg Phe 530 535 540 Ser Ala Gly Gln Pro Ser Leu Leu Arg Glu Thr Ala Leu Ile Trp Leu 545 550 555 560 Asp Val Tyr Gly Gln Thr His Trp Asp Leu Thr Pro Thr Thr Pro Ala 565 570 575 Thr Pro Leu Ser Ala Leu Leu Pro Val Gly Pro Pro Ser His Ala Pro 580 585 590 Ser Val His Leu Ala Ala Ala Thr Lys Ile Arg Phe Pro Ala Leu Glu 595 600 605 Gly Ile His Pro Asn Val Leu Ala Asp Pro Gly Phe Val Pro Tyr Val 610 615 620 Leu Ala Leu Val Val Gly Asp Ala Leu Arg Ala Thr Cys Asn Ala Ala 625 630 635 640 Tyr Leu Pro Arg Pro Ile Glu Phe Ala Leu Arg Val Leu Ala Trp Ala 645 650 655 Arg Asp Phe Gly Leu Gly Tyr Leu Pro Thr Val Glu Gly His Arg Thr 660 665 670 Lys Leu Gly Ala Leu Ile Thr Leu Leu Glu Pro Ala Thr Arg Ala Gly 675 680 685 Val Gly Pro Thr Met Gln Met Ala Asp Asn Ile Glu Gln Leu Leu Arg 690 695 700 Glu Leu Tyr Val Ile Ala Arg Gly Ala Val Glu Gln Leu Arg Pro Ala 705 710 715 720 Val Gln Leu Pro Pro Pro Gln Pro Pro Glu Val Gly Ser Ser Leu Leu 725 730 735 Leu Ile Ser Met Tyr Ala Leu Ala Ala Arg Gly Val Leu Gln Glu Leu 740 745 750 Ala Glu Arg Ala Asp Pro Leu Val Arg Gln Leu Glu Asp Ala Ile Val 755 760 765 Leu Leu Arg Leu His Met Arg Thr Leu Ala Ala Phe Phe Glu Cys Arg 770 775 780 Phe Glu Ser Asp Gly His Arg Leu Tyr Ala Val Val Ala Asp Ala His 785 790 795 800 Glu Arg Leu Gly Pro Trp Arg Pro Glu Ala Met Gly Asp Ala Val Ser 805 810 815 Gln Tyr Cys Gly Met Tyr His Asp Ala Lys Arg Ala Leu Val Ala Ser 820 825 830 Leu Ala Gly Leu Arg Ser Val Val Thr Glu Thr Thr Ala His Leu Gly 835 840 845 Val Cys Asp Glu Leu Ala Ala Gln Val Ser His Glu Gly Asn Val Leu 850 855 860 Ala Val Val Arg Arg Glu Ile His Gly Phe Leu Ala Ile Val Ser Gly 865 870 875 880 Ile His Ala Arg Ala Ser Lys Leu Met Ser Gly Asp Gln Val Pro Gly 885 890 895 Phe Cys Tyr Met Ser Gln Phe Leu Ala Arg Trp Arg Arg Leu Ser Ala 900 905 910 Gly Tyr Gln Ala Ala Arg Ala Ala Thr Gly Pro Glu Arg Val Ala Glu 915 920 925 Phe Val Gln Glu Leu His Asp Thr Trp Lys Gly Leu Gln Thr Glu Arg 930 935 940 Ala Leu Val Val Ala Arg Phe Ala Ser Ser Ala Asp Gln Arg Thr Ala 945 950 955 960 Ala Ile Gln Glu Val Met Ala His Ala Thr Glu Asp Ala Pro Pro Ser 965 970 975 Pro Ala Ala Asp Leu Val Val Leu Thr Asn Arg His Asp Leu Gly Ala 980 985 990 Trp Gly Asp Tyr Ser Leu Gly Pro Leu Gly Gln Pro Thr Val Val Pro 995 1000 1005 Asp Ser Val Asp Leu Ser Pro Gln Gly Leu Ala Ala Thr Leu Ser Met 1010 1015 1020 Asp Trp Leu Leu Ile Asn Glu Leu Leu Gln Val Thr Asp Gly Val Phe 1025 1030 1035 1040 Arg Ala Ser Ala Phe Arg Pro Ser Ala Gly Pro Gly Ala Pro Gly Asp 1045 1050 1055 Leu Glu Ala Gln Asp Ala Gly Gly Ser Thr Pro Glu Pro Thr Thr Pro 1060 1065 1070 Gly Pro Gln Asp Thr Gln Ala Arg Ala Pro Ser Thr Pro Ala Gly Arg 1075 1080 1085 Glu Thr Val Pro Trp Pro Asn Thr Pro Val Glu Asp Asp Glu Met Thr 1090 1095 1100 Pro Gln Glu Thr Pro Pro Val His Pro 1105 1110 <210> SEQ ID NO 52 <211> LENGTH: 3113 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 52 atgtcggaca gcgcgctcca ggtcccggcg cccgccggca tgaccccccc gtccgctcca 60 cccccaaacg gaccgttgca ggtcctgttg gggagcctga ccaacctccg ccggccgccc 120 tccccctcga gcgagccggc gggcagcgcg gacgagccgg cgtttctgtc cgcggccaaa 180 ctgcgcgccg ccacggcggc gtttctgttg tcgggcgcgg cggtcggccc ggcggaggcg 240 cgggcgtgct ggcacccgct cctggagcaa ctgtgcgcct tgcaccgggc ccacgggctt 300 ccggagacgg cgctcttggc cgagaacctc cccgggctgc tggtgcaccg catggcggtg 360 gcgctccccg agacccccga ggcggccttc cgggagatgg acgtcatcaa ggacacggtc 420 ctggcgatca ccggctccga cacgacccac gcgctggagg ccgccgggct gcgcaccacg 480 gcggccctgg ggccggtgcg cgtccgccag tgtgccgtgg agtggatcga ccgctggcgg 540 accgtcaccc agagctgcct ggccatgaac ccgcggacct ccctggaggc cctcggggag 600 atgtcgctta agatgtcccc ggttcccctg gggcaacccg gcgccaacct gaccaccccg 660 gcgtacagcc tgctcttccc gtccccgatc gtgcaagagg gtctccggtt cttggccctg 720 gtcagcaatt gggtgacgct gttctcggcg cacctacagc gcatcgacga cgccgccctg 780 acccccctca cgcgggccct gtttacgctg gcgctggtgg acgagtacct gacgaccccc 840 gatcggggcg cggtggtccc gccgccgctg ctggcccagt ttcagcacac ggtgcgggag 900 atcgacccgg ctatcatgat cccgcccctc gaggccacca aaatggttcg cagtcgcgag 960 gaggtgcgcg tgtcgaccgc cctaagccgc gtcagcccgc ggtcggcgtg cgcgcccccg 1020 gggacgctga tggcgcgcgt gcggacggac gcggccgtct tcgaccccga cgtgccgttt 1080 ctgagtgcgt cggcgctggc catcttccgt ccggccgtca ccggcctgct gcagctaggg 1140 gagccgccct ccgccggggc ccagcaacgg ctcctcgcgc tgctgcagca gacgtgggct 1200 ttggtccaga acagcaactc gccctccgtg gtgatcaaca cgcttaccga cgcggggttc 1260 acgcccgcgc actgcaccca gtacatatcg gccctggagg ggtttttggt ggcgggggtg 1320 cccgcgcgga cgccccccgg ccacgggctc agcgagatcc agcagctctt tgggtgcatt 1380 gcccttgcgg gggcgaacgt gttcgggctc gcgcgggagt acgggcacta cgccggctac 1440 gtgaaaacgt tccggcggat ccagggcgcc agcgagcaca cgcacgggcg gctctgtgag 1500 gcggtcggcc tgtcgggcgg cgtcctcagc cagacgctgg cgcgcatcat gggcccggcc 1560 gtgccgacgg aacacctggc gagcctgcgg cgcacgctcg tgggggagtt tgagacggcc 1620 gagcgccgct tcagcgccgg ccagcccagc ctcctgcgcg agacggcgct catctggctc 1680 gacgtgtacg gccagaccca ctgggacctg acccccacca ccccggccac gccgctgtcc 1740 gcgctgctcc ccgtcgggcc gcctagccac gccccctcgg ttcacctggc cgcggccacc 1800 aagatccgct tcccggccct ggagggcatc caccccaacg tcctcgccga cccggggttc 1860 gtcccttacg tgctggccct ggtggtcggg gacgcgctga gggccacgtg caacgctgcc 1920 tacctgcccc gccccatcga gtttgccctg cgggtgctgg cctgggcgcg cgacttcggc 1980 ctgggctatc tccccaccgt cgaggggcac cgcacaaaat tgggcgcgct gatcaccctc 2040 ctcgaaccgg ccacccgggc cggcgtcggg cccacgatgc agatggccga caacatcgag 2100 cagctgctgc gcgagctgta cgtgatcgcc aggggggccg tcgagcagct gcggcccgcg 2160 gtccagctgc ctccccccca gcctcccgag gtgggcagca gcctgctgct gatcagcatg 2220 tacgccctgg cggcccgggg ggtgctgcag gagctcgccg agcgcgccga ccccctggtc 2280 cgccagctgg aggacgcgat cgtgctgctg cggcttcaca tgcgcacgct cgccgccttc 2340 tttgagtgcc ggttcgagag cgacgggcac cgcctgtacg ccgtggtcgc agacgcccac 2400 gagcgcctgg ggccctggcg ccccgaggcc atgggcgacg cggtgagcca gtactgcggc 2460 atgtatcacg acgccaagcg cgcgctggtc gcgtccctcg cggggctgcg gtccgtcgtc 2520 accgagacga cggcgcacct cggggtatgc gacgagctgg cggcccaggt ctcgcacgag 2580 ggtaacgtgc tggccgtcgt gcggcgggaa atccacgggt tcctggccat cgtgtcgggc 2640 atccacgcca gggcgtcaaa gttgatgtcg ggggaccagg tccccgggtt ctgttacatg 2700 agtcagttcc tggcgcgctg gcggcgcctc tcggccggct atcaggccgc gcgcgcggcc 2760 acgggccccg agcgggtggc cgagttcgtt caggaactcc acgacacgtg gaagggcctg 2820 cagacggagc gcgcgctggt cgtggcgccc ttcgccagct cggccgatca gcgcaccgca 2880 gccattcagg aggtgatggc ccacgcgacc gaggacgcgc cccccagccc ggccgcagac 2940 ctcgtcgtgc tcacgaaccg tcatgacctg ggggcctggg gggactacag cctggggccc 3000 ctgggtcagc cgacggtggt tccggactcc gtggacctgt ctccccaggg gctggccgct 3060 acgctgagca tggactggct attaataaac gagctcctgc aggtcaccga cgg 3113 <210> SEQ ID NO 53 <211> LENGTH: 761 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 53 gcgcccgctc gcggctcagc gcgaggccgc cggggtttac gacgcggtgc ggacctgggg 60 gccagacgcg gaggccgaac cggaccagat ggaaaacacg tatctgctgc ccgacgatga 120 cgccgccatg cccgcgggcg tcgggcttgg cgccaccccc gccgccgaca ccaccgccgc 180 cgcctggccg gccgaaagcc acgccccccg cgccccctcc gaggacgcag attccattta 240 cgagtcggtg agcgaggatg gggggcgcgt ctacgaggag atcccytggg ttcgggtata 300 cgaaaacatc tgccttcgcc ggcaagacgc cggcggggcg gccccgccgg gagacgcccc 360 ggactccccg tacatcgagg cggaaaatcc cctgtacgac tggggcgggt ctgccctctt 420 ctcccctccg ggggccacac gcgccccgga cccgggacta agcctgtcgc ccatgcccgc 480 ccgcccccgg accaacgcgc tggccaacga cggcccgaca aacgtcgccg ccctcagcgc 540 cctgttgacg aagctcaaac gcggccgaca ccagagccat taaaaaaatg cgaccgccgg 600 ccccaccgtc tcggtttccg gcccctttcc ccgtatgtct gttttcaata aaaagtaaca 660 aacagagaaa aaaaaacagc gagttccgca tggtttgtcg tacgcaatta gctgtttatt 720 gttttttttt tggggggggg aagagaaaaa gaaaaaagga g 761 <210> SEQ ID NO 54 <211> LENGTH: 1037 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 54 Met Ser Asp Ser Ala Leu Gln Val Pro Ala Pro Ala Gly Met Thr Pro 5 10 15 Pro Ser Ala Pro Pro Pro Asn Gly Pro Leu Gln Val Leu Leu Gly Ser 20 25 30 Leu Thr Asn Leu Arg Arg Pro Pro Ser Pro Ser Ser Glu Pro Ala Gly 35 40 45 Ser Ala Asp Glu Pro Ala Phe Leu Ser Ala Ala Lys Leu Arg Ala Ala 50 55 60 Thr Ala Ala Phe Leu Leu Ser Gly Ala Ala Val Gly Pro Ala Glu Ala 65 70 75 80 Arg Ala Cys Trp His Pro Leu Leu Glu Gln Leu Cys Ala Leu His Arg 85 90 95 Ala His Gly Leu Pro Glu Thr Ala Leu Leu Ala Glu Asn Leu Pro Gly 100 105 110 Leu Leu Val His Arg Met Ala Val Ala Leu Pro Glu Thr Pro Glu Ala 115 120 125 Ala Phe Arg Glu Met Asp Val Ile Lys Asp Thr Val Leu Ala Ile Thr 130 135 140 Gly Ser Asp Thr Thr His Ala Leu Glu Ala Ala Gly Leu Arg Thr Thr 145 150 155 160 Ala Ala Leu Gly Pro Val Arg Val Arg Gln Cys Ala Val Glu Trp Ile 165 170 175 Asp Arg Trp Arg Thr Val Thr Gln Ser Cys Leu Ala Met Asn Pro Arg 180 185 190 Thr Ser Leu Glu Ala Leu Gly Glu Met Ser Leu Lys Met Ser Pro Val 195 200 205 Pro Leu Gly Gln Pro Gly Ala Asn Leu Thr Thr Pro Ala Tyr Ser Leu 210 215 220 Leu Phe Pro Ser Pro Ile Val Gln Glu Gly Leu Arg Phe Leu Ala Leu 225 230 235 240 Val Ser Asn Trp Val Thr Leu Phe Ser Ala His Leu Gln Arg Ile Asp 245 250 255 Asp Ala Ala Leu Thr Pro Leu Thr Arg Ala Leu Phe Thr Leu Ala Leu 260 265 270 Val Asp Glu Tyr Leu Thr Thr Pro Asp Arg Gly Ala Val Val Pro Pro 275 280 285 Pro Leu Leu Ala Gln Phe Gln His Thr Val Arg Glu Ile Asp Pro Ala 290 295 300 Ile Met Ile Pro Pro Leu Glu Ala Thr Lys Met Val Arg Ser Arg Glu 305 310 315 320 Glu Val Arg Val Ser Thr Ala Leu Ser Arg Val Ser Pro Arg Ser Ala 325 330 335 Cys Ala Pro Pro Gly Thr Leu Met Ala Arg Val Arg Thr Asp Ala Ala 340 345 350 Val Phe Asp Pro Asp Val Pro Phe Leu Ser Ala Ser Ala Leu Ala Ile 355 360 365 Phe Arg Pro Ala Val Thr Gly Leu Leu Gln Leu Gly Glu Pro Pro Ser 370 375 380 Ala Gly Ala Gln Gln Arg Leu Leu Ala Leu Leu Gln Gln Thr Trp Ala 385 390 395 400 Leu Val Gln Asn Ser Asn Ser Pro Ser Val Val Ile Asn Thr Leu Thr 405 410 415 Asp Ala Gly Phe Thr Pro Ala His Cys Thr Gln Tyr Ile Ser Ala Leu 420 425 430 Glu Gly Phe Leu Val Ala Gly Val Pro Ala Arg Thr Pro Pro Gly His 435 440 445 Gly Leu Ser Glu Ile Gln Gln Leu Phe Gly Cys Ile Ala Leu Ala Gly 450 455 460 Ala Asn Val Phe Gly Leu Ala Arg Glu Tyr Gly His Tyr Ala Gly Tyr 465 470 475 480 Val Lys Thr Phe Arg Arg Ile Gln Gly Ala Ser Glu His Thr His Gly 485 490 495 Arg Leu Cys Glu Ala Val Gly Leu Ser Gly Gly Val Leu Ser Gln Thr 500 505 510 Leu Ala Arg Ile Met Gly Pro Ala Val Pro Thr Glu His Leu Ala Ser 515 520 525 Leu Arg Arg Thr Leu Val Gly Glu Phe Glu Thr Ala Glu Arg Arg Phe 530 535 540 Ser Ala Gly Gln Pro Ser Leu Leu Arg Glu Thr Ala Leu Ile Trp Leu 545 550 555 560 Asp Val Tyr Gly Gln Thr His Trp Asp Leu Thr Pro Thr Thr Pro Ala 565 570 575 Thr Pro Leu Ser Ala Leu Leu Pro Val Gly Pro Pro Ser His Ala Pro 580 585 590 Ser Val His Leu Ala Ala Ala Thr Lys Ile Arg Phe Pro Ala Leu Glu 595 600 605 Gly Ile His Pro Asn Val Leu Ala Asp Pro Gly Phe Val Pro Tyr Val 610 615 620 Leu Ala Leu Val Val Gly Asp Ala Leu Arg Ala Thr Cys Asn Ala Ala 625 630 635 640 Tyr Leu Pro Arg Pro Ile Glu Phe Ala Leu Arg Val Leu Ala Trp Ala 645 650 655 Arg Asp Phe Gly Leu Gly Tyr Leu Pro Thr Val Glu Gly His Arg Thr 660 665 670 Lys Leu Gly Ala Leu Ile Thr Leu Leu Glu Pro Ala Thr Arg Ala Gly 675 680 685 Val Gly Pro Thr Met Gln Met Ala Asp Asn Ile Glu Gln Leu Leu Arg 690 695 700 Glu Leu Tyr Val Ile Ala Arg Gly Ala Val Glu Gln Leu Arg Pro Ala 705 710 715 720 Val Gln Leu Pro Pro Pro Gln Pro Pro Glu Val Gly Ser Ser Leu Leu 725 730 735 Leu Ile Ser Met Tyr Ala Leu Ala Ala Arg Gly Val Leu Gln Glu Leu 740 745 750 Ala Glu Arg Ala Asp Pro Leu Val Arg Gln Leu Glu Asp Ala Ile Val 755 760 765 Leu Leu Arg Leu His Met Arg Thr Leu Ala Ala Phe Phe Glu Cys Arg 770 775 780 Phe Glu Ser Asp Gly His Arg Leu Tyr Ala Val Val Ala Asp Ala His 785 790 795 800 Glu Arg Leu Gly Pro Trp Arg Pro Glu Ala Met Gly Asp Ala Val Ser 805 810 815 Gln Tyr Cys Gly Met Tyr His Asp Ala Lys Arg Ala Leu Val Ala Ser 820 825 830 Leu Ala Gly Leu Arg Ser Val Val Thr Glu Thr Thr Ala His Leu Gly 835 840 845 Val Cys Asp Glu Leu Ala Ala Gln Val Ser His Glu Gly Asn Val Leu 850 855 860 Ala Val Val Arg Arg Glu Ile His Gly Phe Leu Ala Ile Val Ser Gly 865 870 875 880 Ile His Ala Arg Ala Ser Lys Leu Met Ser Gly Asp Gln Val Pro Gly 885 890 895 Phe Cys Tyr Met Ser Gln Phe Leu Ala Arg Trp Arg Arg Leu Ser Ala 900 905 910 Gly Tyr Gln Ala Ala Arg Ala Ala Thr Gly Pro Glu Arg Val Ala Glu 915 920 925 Phe Val Gln Glu Leu His Asp Thr Trp Lys Gly Leu Gln Thr Glu Arg 930 935 940 Ala Leu Val Val Ala Pro Phe Ala Ser Ser Ala Asp Gln Arg Thr Ala 945 950 955 960 Ala Ile Gln Glu Val Met Ala His Ala Thr Glu Asp Ala Pro Pro Ser 965 970 975 Pro Ala Ala Asp Leu Val Val Leu Thr Asn Arg His Asp Leu Gly Ala 980 985 990 Trp Gly Asp Tyr Ser Leu Gly Pro Leu Gly Gln Pro Thr Val Val Pro 995 1000 1005 Asp Ser Val Asp Leu Ser Pro Gln Gly Leu Ala Ala Thr Leu Ser Met 1010 1015 1020 Asp Trp Leu Leu Ile Asn Glu Leu Leu Gln Val Thr Asp 1025 1030 1035 <210> SEQ ID NO 55 <211> LENGTH: 193 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 55 Arg Pro Leu Ala Ala Gln Arg Glu Ala Ala Gly Val Tyr Asp Ala Val 5 10 15 Arg Thr Trp Gly Pro Asp Ala Glu Ala Glu Pro Asp Gln Met Glu Asn 20 25 30 Thr Tyr Leu Leu Pro Asp Asp Asp Ala Ala Met Pro Ala Gly Val Gly 35 40 45 Leu Gly Ala Thr Pro Ala Ala Asp Thr Thr Ala Ala Ala Trp Pro Ala 50 55 60 Glu Ser His Ala Pro Arg Ala Pro Ser Glu Asp Ala Asp Ser Ile Tyr 65 70 75 80 Glu Ser Val Ser Glu Asp Gly Gly Arg Val Tyr Glu Glu Ile Pro Trp 85 90 95 Val Arg Val Tyr Glu Asn Ile Cys Leu Arg Arg Gln Asp Ala Gly Gly 100 105 110 Ala Ala Pro Pro Gly Asp Ala Pro Asp Ser Pro Tyr Ile Glu Ala Glu 115 120 125 Asn Pro Leu Tyr Asp Trp Gly Gly Ser Ala Leu Phe Ser Pro Pro Gly 130 135 140 Ala Thr Arg Ala Pro Asp Pro Gly Leu Ser Leu Ser Pro Met Pro Ala 145 150 155 160 Arg Pro Arg Thr Asn Ala Leu Ala Asn Asp Gly Pro Thr Asn Val Ala 165 170 175 Ala Leu Ser Ala Leu Leu Thr Lys Leu Lys Arg Gly Arg His Gln Ser 180 185 190 His <210> SEQ ID NO 56 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 56 Ser Pro Asn Thr Asp Val Arg Met Tyr Ser Gly Lys Arg Asn Gly 5 10 15 <210> SEQ ID NO 57 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 57 Tyr Leu Ala Ala Pro Thr Gly Ile Pro Pro Ala Phe Phe Pro Ile 5 10 15 <210> SEQ ID NO 58 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 58 Gly Val Ala Ala Ala Thr Pro Arg Pro Asp Pro Glu Asp Gly Ala 5 10 15 <210> SEQ ID NO 59 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 59 Glu Glu Ile Pro Trp Val Arg Val Tyr Glu Asn Ile Cys Pro Arg 5 10 15 <210> SEQ ID NO 60 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 60 Pro Gly Asp Ala Pro Asp Ser Pro Tyr Ile Glu Ala Glu Asn Pro 5 10 15 <210> SEQ ID NO 61 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 61 Pro Asp Ser Pro Tyr Ile Glu Ala Glu Asn Pro Leu Tyr Asp Trp 5 10 15 <210> SEQ ID NO 62 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 62 Glu Asn Pro Leu Tyr Asp Trp Gly Gly Ser Ala Leu Phe Ser Pro 5 10 15 <210> SEQ ID NO 63 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 63 Ala Ile Asp Tyr Val His Cys Glu Gly Ile Ile His Arg Asp Ile 5 10 15 <210> SEQ ID NO 64 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 64 Ala Phe Pro Val Ala Leu His Ala Val Asp Ala Pro Ser Gln Phe 5 10 15 <210> SEQ ID NO 65 <211> LENGTH: 3429 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 65 atggccaacc gccctgccgc atccgccctc gccggagcgc ggtctccgtc cgaacgacag 60 gaaccccggg agcccgaggt cgccccccct ggcggcgacc acgtgttttg caggaaagtc 120 agcggcgtga tggtgctttc cagcgatccc cccggccccg cggcctaccg cattagcgac 180 agcagctttg ttcaatgcgg ctccaactgc agtatgataa tcgacggaga cgtggcgcgc 240 ggtcatttgc gtgacctcga gggcgctacg tccaccggcg ccttcgtcgc gatctcaaac 300 gtcgcagccg gcggggatgg ccgaaccgcc gtcgtggcgc tcggcggaac ctcgggcccg 360 tccgcgacta catccgtggg gacccagacg tccggggagt tcctccacgg gaacccaagg 420 acccccgaac cccaaggacc ccaggctgtc cccccgcccc ctcctccccc ctttccatgg 480 ggccacgagt gctgcgcccg tcgcgatgcc aggggcggcg ccgagaagga cgtcggggcc 540 gcggagtcat ggtcagacgg cccgtcgtcc gactccgaaa cggaggactc ggactcctcg 600 gacgaggata cgggttcgga gacgctgtct cgatcctctt cgatctgggc cgcaggggcg 660 actgacgacg atgacagcga ctccgactcg cggtcggacg actccgtgca gcccgacgtt 720 gtcgttcgtc gcagatggag cgacggcccc gcccccgtgg cctttcccaa gccccggcgc 780 cccggcgact cccccggaaa ccccggcctg ggcgccggca ccgggccggg ctccgcgacg 840 gacccgcgcg cgtcggccga ctccgattcc gcggcccacg ccgccgcacc ccaggcggac 900 gtggcgccgg ttctggacag ccagcccact gtgggaacgg accccggcta cccagtcccc 960 ctagaactca cgcccgagaa cgcggaggcg gtggcgcggt ttctggggga cgccgtcgac 1020 cgcgagcccg cgctcatgct ggagtacttc tgtcggtgcg cccgcgagga gagcaagcgc 1080 gtgcccccac gaaccttcgg cagcgccccc cgcctcacgg aggacgactt tgggctcctg 1140 aactacgcgc tcgctgagat gcgacgcctg tgcctggacc ttcccccggt cccccccaac 1200 gcatacacgc cctatcatct gagggagtat gcgacgcggc tggttaacgg gttcaaaccc 1260 ctggtgcggc ggtccgcccg cctgtatcgc atcctggggg ttctggtcca cctgcgcatc 1320 cgtacccggg aggcctcctt tgaggaatgg atgcgctcca aggaggtgga cctggacttc 1380 gggctgacgg aaaggcttcg cgaacacgag gcccagctaa tgatcctggc ccaggccctg 1440 aacccctacg actgtctgat ccacagcacc ccgaacacgc tcgtcgagcg ggggctgcag 1500 tcggcgctga agtacgaaga gttttacctc aagcgcttcg gcgggcacta catggagtcc 1560 gtcttccaga tgtacacccg catcgccggg tttctggcgt gccgggcgac ccgcggcatg 1620 cgccacatcg ccctggggcg acaggggtcg tggtgggaaa tgttcaagtt ctttttccac 1680 cgcctctacg accaccagat cgtgccgtcc acccccgcca tgctgaacct cggaacccgc 1740 aactactaca cgtccagctg ctacctggta aacccccagg ccaccactaa ccaggccacc 1800 ctccgggcca tcaccggcaa cgtgagcgcc atcctcgccc gcaacggggg catcgggctg 1860 tgcatgcagg cgttcaacga cgccagcccc ggcaccgcca gcatcatgcc ggccctgaag 1920 gtcctcgact ccctggtggc ggcgcacaac aaacagagca cgcgccccac cggggcgtgc 1980 gtgtacctgg aaccctggca cagcgacgtt cgggccgtgc tcagaatgaa gggcgtcctc 2040 gccggcgagg aggcccagcg ctgcgacaac atcttcagcg ccctctggat gccggacctg 2100 ttcttcaagc gcctgatccg ccacctcgac ggcgagaaaa acgtcacctg gtccctgttc 2160 gaccgggaca ccagcatgtc gctcgccgac tttcacggcg aggagttcga gaagctgtac 2220 gagcacctcg aggccatggg gttcggcgaa acgatcccca tccaggacct ggcgtacgcc 2280 atcgtgcgca gcgcggccac caccggaagc cccttcatca tgtttaagga cgcggtaaac 2340 cgccactaca tctacgacac gcaaggggcg gccatcgccg gctccaacct ctgcaccgag 2400 atcgtccacc cggcctccaa gcgatccagt ggggtctgca acctgggaag cgtgaatctg 2460 gcccgatgcg tctccaggca gacgtttgac tttgggcggc tccgcgacgc cgtgcaggcg 2520 tgcgtgctga tggtgaacat catgatcgac agcacgctac aacccacgcc ccagtgcacc 2580 cgcggcaacg acaacctgcg gtccatgggc attggcatgc agggcctgca cacggcgtgc 2640 ctcaagatgg gcctggatct ggagtcggcc gagttccggg acctgaacac acacatcgcc 2700 gaggtgatgc tgctcgcggc catgaagacc agtaacgcgc tgtgcgttcg cggggcgcgt 2760 cccttcagcc actttaagcg cagcatgtac cgggccggcc gctttcactg ggagcgcttt 2820 tcgaacgcca gcccgcggta cgagggcgag tgggagatgc tacgccagag catgatgaaa 2880 cacggcctgc gcaacagcca gttcatcgcg ctcatgccca ccgccgcctc ggcccagatc 2940 tcggacgtca gcgagggctt tgcccccctg ttcaccaacc tgttcagcaa ggtgaccagg 3000 gacggcgaga cgctgcgccc caacacgctc ttgctgaagg aactcgagcg cacgttcggc 3060 gggaagcggc tcctggacgc gatggacggg ctcgaggcca agcagtggtc tgtggcccag 3120 gccctgcctt gcctggaccc cgcccacccc ctccggcggt tcaagacggc cttcgactac 3180 gaccaggaac tgctgatcga cctgtgtgca gaccgcgccc cctatgttga tcacagccaa 3240 tccatgactc tgtatgtcac agagaaggcg gacgggacgc tccccgcctc caccctggtc 3300 cgccttctcg tccacgcata taagcgcggc ctgaagacgg ggatgtacta ctgcaaggtt 3360 cgcaaggcga ccaacagcgg ggtgttcgcc ggcgacgaca acatcgtctg cacaagctgc 3420 gcgctgtaa 3429 <210> SEQ ID NO 66 <211> LENGTH: 825 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 66 ggaaagtcag cggcgtgatg gtgctttcca gcgatccccc cggccccgcg gcctaccgca 60 ttagcgacag cagctttgtt caatgcggct ccaactgcag tatgataatc gacggagacg 120 tggcgcgcgg tcatttgcgt gacctcgagg gcgctacgtc caccggcgcc ttcgtcgcga 180 tctcaaacgt cgcagccggc ggggatggcc gaaccgccgt cgtggcgctc ggcggaacct 240 cgggcccgtc cgcgactaca tccgtgggga cccagacgtc cggggagttc ctccacggga 300 acccaaggac ccccgaaccc caaggacccc aggctgtccc cccgccccct cctcccccct 360 ttccatgggg ccacgagtgc tgcgcccgtc gcgatgccag gggcggcgcc gagaaggacg 420 tcggggccgc ggagtcatgg tcagacggcc cgtcgtccga ctccgaaacg gaggactcgg 480 actcctcgga cgaggatacg ggctcgggtt cggagacgct gtctcgatcc tcttcgatct 540 gggccgcagg ggcgactgac gacgatgaca gcgactccga ctcgcggtcg gacgactccg 600 tgcagcccga cgttgtcgtt cgtcgcagat ggagcgacgg ccccgccccc gtggcctttc 660 ccaagccccg gcgccccggc gactcccccg gaaaccccgg cctgggcgcc ggcaccgggc 720 cgggctccgc gacggacccg cgcgcgtcgg ccgactccga ttccgcggcc cacgccgccg 780 caccccaggc ggacgtggcg ccggttctgg acagccagcc cactg 825 <210> SEQ ID NO 67 <211> LENGTH: 678 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 67 atggccaacc gccctgccgc atccgccctc gccggagcgc ggtctccgtc cgaacgacag 60 gaaccccggg agcccgaggt cgccccccct ggcggcgacc acgtgttttg caggaaagtc 120 agcggcgtga tggtgctttc cagcgatccc cccggccccg cggcctaccg cattagcgac 180 agcagctttg ttcaatgcgg ctccaactgc agtatgataa tcgacggaga cgtggcgcgc 240 ggtcatttgc gtgacctcga gggcgctacg tccaccggcg ccttcgtcgc gatctcaaac 300 gtcgcagccg gcggggatgg ccgaaccgcc gtcgtggcgc tcggcggaac ctcgggcccg 360 tccgcgacta catccgtggg gacccagacg tccggggagt tcctccacgg gaacccaagg 420 acccccgaac cccaaggacc ccaggctgtc cccccgcccc ctcctccccc ctttccatgg 480 ggccacgagt gctgcgcccg tcgcgatgcc aggggcggcg ccgagaagga cgtcggggcc 540 gcggagtcat ggtcagacgg cccgtcgtcc gactccgaaa cggaggactc ggactcctcg 600 gacgaggata cgggctcggg ttcggagacg ctgtctcgat cctcttcgat ctgggccgca 660 ggggcgactg acgacgat 678 <210> SEQ ID NO 68 <211> LENGTH: 313 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 68 gacgaggggt cggaatccaa aggacgcaga ccacctttgg ttacggaccc ctttctcccc 60 cccttccgaa caaaaagcag cgggcggggg gccggggtga gggagggaca cgggggacac 120 ggcacggggg tcccgcctca cgccccgcgc cctctaaatc ccccccgttt ctttgtcaag 180 cagcccgccg ccccgcacgc ctgggggatg ctcaacgaca tgcagtggct cgccagcagc 240 gactcggagg aggagaccga ggtgggaatc tctgacgacg accttcaccg cgactccacc 300 tccgaggcgg gca 313 <210> SEQ ID NO 69 <211> LENGTH: 467 <212> TYPE: DNA <213> ORGANISM: HSV-2 <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 39,322,332,368,369 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 69 cagggcagcc ccacgcccgc cgacgcgcag ggatcctgnt gggggtgggc ccgtgggtga 60 ggaggaagcg gaagcgggag gggggggcga cgtgtgcgcc gtgtgcacgg acgagatcgc 120 cccgcccctg cgctgccaga gttttccctg cctgcacccc ttctgcatcc cgtgcatgaa 180 gacctggatt ccgttgcgca acacgtgtcc cctgtgcaac accccggtgg cgtacctgat 240 agtgggcgtg accgccagcg ggtcgttcag caccatcccg atagtgaacg acccccggac 300 ccgcgtggag gccgaggcgg cngtgcgggt cnggcacggc cgtggacttt atctggacgg 360 gcaacccnng gacggccccg cgctccctgt cgctgggggg acacacggtc cgcgccctgt 420 cgcccacccc cccgtggccc ggcacggacg acgaggacga tgacctc 467 <210> SEQ ID NO 70 <211> LENGTH: 204 <212> TYPE: DNA <213> ORGANISM: HSV-2 <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 78,79,120,121,124,125 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 70 gcccctattg gtcccctggg cttcctagta tgctaatgaa tttctccccg ggggcgggca 60 ccactcaggg ccgcgcgnng ggccgcgggg gactcccatc tgcgtcggcg gggggcggcn 120 natnntaatg gggttcttgg agtacacccg gttggtcccc ggggacggcc cgccccgaga 180 gggggattcc ctccctccgc cccc 204 <210> SEQ ID NO 71 <211> LENGTH: 474 <212> TYPE: DNA <213> ORGANISM: HSV-2 <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 7,43,56,339,424,431,451,468,474 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 71 ccccggnccg cttaagcggt cgggggaccc ccgtgggccg tgngccgccc cccgancctc 60 tgggggggcg agggaggcag ggaggagccc gagagcgggg gacagggggg gagacgaggg 120 gtcggaatcc aaaggacgca gaccaccttt ggttacggac ccctttctcc cccccttccg 180 aacaaaaagc agcgggcggg gggccggggt gagggaggga cacgggggac acggcacggg 240 ggtcccgcct cacgccccgc gccctctaaa tcccccccgt ttctttgtca agcagcccgc 300 cgccccgcac gcctggggga tgctcaacga catgcagtng ctcgccagca gcgactcgga 360 ggaggagacc gaggtgggaa tctctgacga cgaccttcac cgcgactcca cctccgaggc 420 gggncagcac nggacacgga gatgttcgag ncgggcctga tggacgcngc cacn 474 <210> SEQ ID NO 72 <211> LENGTH: 350 <212> TYPE: DNA <213> ORGANISM: HSV-2 <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 107,148,185,187,305,312,313 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 72 ggggtcggaa tccaaaggac gcagaccacc tttggttacg gacccctttc tccccccctt 60 ccgaacaaaa agcagcgggc ggggggccgg ggtgagggag ggacacnggg ggacacggca 120 cgggggtccc gcctcacgcc ccgcgccntc taaatccccc ccgtttcttt gtcaagcagc 180 ccgcngnccc gcacgcctgg gggatgctca acgacatgca gtggctcgcc agcagcgact 240 cggaggagga gaccgaggtg ggaatctctg acgacgacct tcaccgcgac tccacctccg 300 aggcngggca gnncggacac ggagatgttc gaggcgggct tgatggacgc 350 <210> SEQ ID NO 73 <211> LENGTH: 312 <212> TYPE: DNA <213> ORGANISM: HSV-2 <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: 21,32,39,66,306 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 73 gccccacgcc cgccgacgcg nagggatcct gngggggtng gcccgtgggt gaggaggaag 60 cggaancggg aggggggggc gacgtgtgcg ccgtgtgcac ggacgagatc gccccgcccc 120 tgcgctgcca gagttttccc tgcctgcacc ccttctgcat cccgtgcatg aagacctgga 180 ttccgttgcg caacacgtgt cccctgtgca acaccccggt ggcgtacctg atagtgggcg 240 tgaccgccag cgggtcgttc agcaccatcc cgatagtgaa cgacccccgg acccgcgtgg 300 aggccngagg cg 312 <210> SEQ ID NO 74 <211> LENGTH: 274 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 74 Lys Val Ser Gly Val Met Val Leu Ser Ser Asp Pro Pro Gly Pro Ala 5 10 15 Ala Tyr Arg Ile Ser Asp Ser Ser Phe Val Gln Cys Gly Ser Asn Cys 20 25 30 Ser Met Ile Ile Asp Gly Asp Val Ala Arg Gly His Leu Arg Asp Leu 35 40 45 Glu Gly Ala Thr Ser Thr Gly Ala Phe Val Ala Ile Ser Asn Val Ala 50 55 60 Ala Gly Gly Asp Gly Arg Thr Ala Val Val Ala Leu Gly Gly Thr Ser 65 70 75 80 Gly Pro Ser Ala Thr Thr Ser Val Gly Thr Gln Thr Ser Gly Glu Phe 85 90 95 Leu His Gly Asn Pro Arg Thr Pro Glu Pro Gln Gly Pro Gln Ala Val 100 105 110 Pro Pro Pro Pro Pro Pro Pro Phe Pro Trp Gly His Glu Cys Cys Ala 115 120 125 Arg Arg Asp Ala Arg Gly Gly Ala Glu Lys Asp Val Gly Ala Ala Glu 130 135 140 Ser Trp Ser Asp Gly Pro Ser Ser Asp Ser Glu Thr Glu Asp Ser Asp 145 150 155 160 Ser Ser Asp Glu Asp Thr Gly Ser Gly Ser Glu Thr Leu Ser Arg Ser 165 170 175 Ser Ser Ile Trp Ala Ala Gly Ala Thr Asp Asp Asp Asp Ser Asp Ser 180 185 190 Asp Ser Arg Ser Asp Asp Ser Val Gln Pro Asp Val Val Val Arg Arg 195 200 205 Arg Trp Ser Asp Gly Pro Ala Pro Val Ala Phe Pro Lys Pro Arg Arg 210 215 220 Pro Gly Asp Ser Pro Gly Asn Pro Gly Leu Gly Ala Gly Thr Gly Pro 225 230 235 240 Gly Ser Ala Thr Asp Pro Arg Ala Ser Ala Asp Ser Asp Ser Ala Ala 245 250 255 His Ala Ala Ala Pro Gln Ala Asp Val Ala Pro Val Leu Asp Ser Gln 260 265 270 Pro Thr <210> SEQ ID NO 75 <211> LENGTH: 226 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 75 Met Ala Asn Arg Pro Ala Ala Ser Ala Leu Ala Gly Ala Arg Ser Pro 5 10 15 Ser Glu Arg Gln Glu Pro Arg Glu Pro Glu Val Ala Pro Pro Gly Gly 20 25 30 Asp His Val Phe Cys Arg Lys Val Ser Gly Val Met Val Leu Ser Ser 35 40 45 Asp Pro Pro Gly Pro Ala Ala Tyr Arg Ile Ser Asp Ser Ser Phe Val 50 55 60 Gln Cys Gly Ser Asn Cys Ser Met Ile Ile Asp Gly Asp Val Ala Arg 65 70 75 80 Gly His Leu Arg Asp Leu Glu Gly Ala Thr Ser Thr Gly Ala Phe Val 85 90 95 Ala Ile Ser Asn Val Ala Ala Gly Gly Asp Gly Arg Thr Ala Val Val 100 105 110 Ala Leu Gly Gly Thr Ser Gly Pro Ser Ala Thr Thr Ser Val Gly Thr 115 120 125 Gln Thr Ser Gly Glu Phe Leu His Gly Asn Pro Arg Thr Pro Glu Pro 130 135 140 Gln Gly Pro Gln Ala Val Pro Pro Pro Pro Pro Pro Pro Phe Pro Trp 145 150 155 160 Gly His Glu Cys Cys Ala Arg Arg Asp Ala Arg Gly Gly Ala Glu Lys 165 170 175 Asp Val Gly Ala Ala Glu Ser Trp Ser Asp Gly Pro Ser Ser Asp Ser 180 185 190 Glu Thr Glu Asp Ser Asp Ser Ser Asp Glu Asp Thr Gly Ser Gly Ser 195 200 205 Glu Thr Leu Ser Arg Ser Ser Ser Ile Trp Ala Ala Gly Ala Thr Asp 210 215 220 Asp Asp 225 <210> SEQ ID NO 76 <211> LENGTH: 4125 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 76 atggccgctc ctgcccgcga ccccccgggt taccggtacg ccgcggccat cctgcccacc 60 ggctccatcc tgagtacgat cgaggtggcg tcccaccgca gactctttga ttttttcgcc 120 gccgtgcgct ccgacgaaaa cagcctgtat gacgtagagt ttgacgccct gctggggtcc 180 tactgcaaca ccctgtcgct cgtgcgcttt ctggagctcg gcctgtccgt ggcgtgcgtg 240 tgcaccaagt tcccggagct ggcttacatg aacgaagggc gtgtgcagtt cgaggtccac 300 cagcccctca tcgcccgcga cggcccgcac cccgtcgagc agcccgtgca taattacatg 360 acgaaggtca tcgaccgccg ggccctgaac gccgccttca gcctggccac cgaggccatt 420 gccctgctca cgggggaggc cctggacggg acgggcatta gcctgcatcg ccagctgcgc 480 gccatccagc agctcgcgcg caacgtccag gccgtcctgg gggcgtttga gcgcggcacg 540 gccgaccaga tgctgcacgt gctgttggag aaggcgcctc ccctggccct gctgttgccc 600 atgcaacgat atctcgacaa cgggcgcctg gcgaccaggg ttgcccgggc gaccctggtc 660 gccgagctga agcggagctt ttgcgacacg agcttcttcc tgggcaaggc gggccatcgc 720 cgcgaggcca tcgaggcctg gctcgtggac ctgaccacgg cgacgcagcc gtccgtggcc 780 gtgccccgcc tgacgcacgc cgacacgcgc gggcggccgg tcgacggggt gctggtcacc 840 accgccgcca tcaaacagcg cctcctgcag tccttcctga aggtggagga caccgaggcc 900 gacgtgccgg tgacctacgg cgagatggtc ttgaacgggg ccaacctcgt cacggcgctg 960 gtgatgggca aggccgtgcg gagcctggac gacgtgggcc gccacctgct ggatatgcag 1020 gaggagcaac tcgaggcgaa ccgggagacg ctggatgaac tcgaaagcgc cccccagaca 1080 acgcgcgtgc gcgcggatct ggtggccata ggcgacaggc tggtcttcct ggaggccctg 1140 gagagacgca tctacgccgc caccaacgtg ccctaccccc tggtgggcgc catggacctg 1200 acgttcgtcc tgcccctggg gctgttcaac ccggccatgg agcgcttcgc cgcgcacgcc 1260 ggggacctgg tgcccgcccc cggccacccg gagccccgcg cgttccctcc ccggcagctg 1320 tttttttggg gaaaggacca ccaggttctg cggctgtcca tggagaacgc ggtcgggacc 1380 gtgtgtcatc cttcgctcat gaacatcgac gcggccgtcg ggggcgtgaa ccacgacccc 1440 gtcgaggccg cgaatccgta cggggcgtac gtcgcggccc cggccggccc cggcgcggac 1500 atgcagcagc gttttctgaa cgcctggcgg cagcgcctcg cccacggccg ggtccggtgg 1560 gtcgccgagt gccagatgac cgcggagcag ttcatgcagc ccgacaacgc caacctggct 1620 ctggagctgc accccgcgtt cgacttcttc gcgggcgtgg ccgacgtcga gcttcccggc 1680 ggcgaagtcc ccccggccgg tccgggggcg atccaggcca cctggcgcgt ggtcaacggc 1740 aacctgcccc tggcgctgtg tccggtggcg tttcgtgacg cccggggcct ggagctcggc 1800 gttggccgcc acgccatggc gccggctacc atagccgccg tccgcggggc gttcgaggac 1860 cgcagctacc cggcggtgtt ttacctgctg caagccgcga ttcacggcaa cgagcacgtg 1920 ttctgcgccc tggcgcggct cgtgactcag tgcatcacca gctactggaa caacacgcga 1980 tgcgcggcgt tcgtgaacga ctactcgctg gtctcgtaca tcgtgaccta cctcgggggc 2040 gacctccccg aggagtgcat ggccgtgtat cgggacctgg tggcccacgt cgaggccctg 2100 gcccagctgg tggacgactt taccctgccg ggcccggagc tgggcgggca ggctcaggcc 2160 gagctgaatc acctgatgcg cgacccggcg ctgctgccgc ccctcgtgtg ggactgcgac 2220 ggccttatgc gacacgcggc cctggaccgc caccgagact gccggattga cgcggggggg 2280 cacgagcccg tctacgcggc ggcgtgcaac gtggcgacgg ccgactttaa ccgcaacgac 2340 ggccggctgc tgcacaacac ccaggcccgc gcggccgacg ccgccgacga ccggccgcac 2400 cggccggccg actggaccgt ccaccacaaa atctactatt acgtgctggt gccggccttc 2460 tcgcgggggc gctgctgcac cgcgggggtc cgcttcgacc gcgtgtacgc cacgctgcag 2520 aacatggtgg tcccggagat cgcccccggt gaggagtgcc cgagcgatcc cgtgaccgac 2580 cccgcccacc cgctgcatcc cgccaatctg gtggccaaca cggtcaagcg catgttccac 2640 aacgggcgcg tcgtcgtcga cgggcccgcc atgctcacgc tgcaggtgct ggcgcacaac 2700 atggccgagc gcacgacggc gctgctgtgc tccgcggcgc ccgacgcggg cgccaacacc 2760 gcgtcgacgg ccaacatgcg catcttcgac ggggcgctgc acgccggcgt gctgctcatg 2820 gccccccagc acctggacca caccatccaa aatggcgaat acttctacgt cctgcccgtc 2880 cacgcgctgt ttgcgggcgc cgaccacgtg gccaacgcgc ccaacttccc cccggccctg 2940 cgcgacctgg cgcgcgacgt ccccctggtc cccccggccc tgggggccaa ctacttctcc 3000 tccatccgcc agcccgtggt gcagcacgcc cgcgagagcg cggcggggga gaacgcgctg 3060 acctacgcgc tcatggcggg gtacttcaag atgagccccg tggccctgta tcaccagctc 3120 aagacgggcc tccaccccgg gttcgggttc accgtcgtgc ggcaggaccg cttcgtgacc 3180 gagaacgtgc tgttttccga gcgcgcgtcg gaggcgtact ttctgggcca gctccaggtg 3240 gcccgccacg aaacgggcgg gggggtcaac ttcacgctca cccagccgcg cggaaacgtg 3300 gacctgggtg tgggctacac cgccgtcgcg gccacgggca ccgtccgcaa ccccgttacg 3360 gacatgggca acctccccca aaacttttac ctcggccgcg gggccccccc gctgctagac 3420 aacgcggccg ccgtgtacct gcgcaacgcg gtcgtggcgg gaaaccggct ggggccggcc 3480 cagcccctcc cggtctttgg ctgcgcccag gtgccgcggc gcgccggcat ggaccacggg 3540 caggatgccg tgtgtgagtt catcgccacc cccgtggcca cggacatcaa ctactttcgc 3600 cggccctgca acccgcgggg acgcgcggcc ggcggcgtgt acgcggggga caaggagggg 3660 gacgtcatag ccctcatgta cgaccacggc cagagcgacc cggcgcggcc cttcgcggcc 3720 acggccaacc cgtgggcgtc gcagcggttc tcgtacgggg acctgctgta caacggggcc 3780 tatcacctca acggggcctc gcccgtcctc agcccctgct tcaagttctt caccgcggcc 3840 gacatcacgg ccaaacatcg ctgcctggag cgtctcatcg tggaaacggg atcggcggta 3900 tccacggcca ccgctgccag cgacgtgcag tttaagcgcc cgccggggtg ccgcgagctc 3960 gtggaagacc cgtgcggcct gtttcaggaa gcctacccga tcacctgcgc cagcgacccc 4020 gccctgctac gcagcgcccg cgatggggag gcccacgcgc gagagaccca ctttacgcag 4080 tatctcatct acgacgcctc cccgctaaag ggcctgtctc tgtaa 4125 <210> SEQ ID NO 77 <211> LENGTH: 8108 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 77 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 60 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 120 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 180 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 240 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 300 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 360 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 420 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 480 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 540 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 600 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 660 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 720 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 780 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 840 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 900 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 960 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1020 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1080 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1140 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1200 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1260 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1320 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1380 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1440 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1500 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1560 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1620 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1680 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 1740 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 1800 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 1860 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 1920 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 1980 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc 2040 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 2100 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt 2160 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa 2220 cagctatgac catgattacg ccaagctttt gcgatcaata aatggatcac aaccagtatc 2280 tcttaacgat gttcttcgca gatgatgatt cattttttaa gtatttggct agtcaagatg 2340 atgaaatctt cattatctga tatattgcaa atcactcaat atctagactt tctgttatta 2400 ttattgatcc aatcaaaaaa taaattagaa gccgtgggtc attgttatga atctctttca 2460 gaggaataca gacaattgac aaaattcaca gactttcaag attttaaaaa actgtttaac 2520 aaggtcccta ttgttacaga tggaagggtc aaacttaata aaggatattt gttcgacttt 2580 gtgattagtt tgatgcgatt caaaaaagaa tcctctctag ctaccaccgc aatagatcct 2640 gttagataca tagatcctcg tcgcaatatc gcattttcta acgtgatgga tatattaaag 2700 tcgaataaag tgaacaataa ttaattcttt attgtcatca tgaacggcgg acatattcag 2760 ttgataatcg gccccatgtt ttcaggtaaa agtacagaat taattagacg agttagacgt 2820 tatcaaatag ctcaatataa atgcgtgact ataaaatatt ctaacgataa tagatacgga 2880 acgggactat ggacgcatga taagaataat tttgaagcat tggaagcaac taaactatgt 2940 gatctcttgg aatcaattac agatttctcc gtgataggta tcgatgaagg acagttcttt 3000 ccagacattg ttgaattccg agcttggctg caggtcgggg atcccccctg cccggttatt 3060 attatttttg acaccagacc aactggtaat ggtagcgacc ggcgctcagc tggaattccg 3120 ccgatactga cgggctccag gagtcgtcgc caccaatccc catatggaaa ccgtcgatat 3180 tcagccatgt gccttcttcc gcgtgcagca gatggcgatg gctggtttcc atcagttgct 3240 gttgactgta gcggctgatg ttgaactgga agtcgccgcg ccactggtgt gggccataat 3300 tcaattcgcg cgtcccgcag cgcagaccgt tttcgctcgg gaagacgtac ggggtataca 3360 tgtctgacaa tggcagatcc cagcggtcaa aacaggcggc agtaaggcgg tcgggatagt 3420 tttcttgcgg ccctaatccg agccagttta cccgctctgc tacctgcgcc agctggcagt 3480 tcaggccaat ccgcgccgga tgcggtgtat cgctcgccac ttcaacatca acggtaatcg 3540 ccatttgacc actaccatca atccggtagg ttttccggct gataaataag gttttcccct 3600 gatgctgcca cgcgtgagcg gtcgtaatca gcaccgcatc agcaagtgta tctgccgtgc 3660 actgcaacaa cgctgcttcg gcctggtaat ggcccgccgc cttccagcgt tcgacccagg 3720 cgttagggtc aatgcgggtc gcttcactta cgccaatgtc gttatccagc ggtgcacggg 3780 tgaactgatc gcgcagcggc gtcagcagtt gttttttatc gccaatccac atctgtgaaa 3840 gaaagcctga ctggcggtta aattgccaac gcttattacc cagctcgatg caaaaatcca 3900 tttcgctggt ggtcagatgc gggatggcgt gggacgcggc ggggagcgtc acactgaggt 3960 tttccgccag acgccactgc tgccaggcgc tgatgtgccc ggcttctgac catgcggtcg 4020 cgttcggttg cactacgcgt actgtgagcc agagttgccc ggcgctctcc ggctgcggta 4080 gttcaggcag ttcaatcaac tgtttacctt gtggagcgac atccagaggc acttcaccgc 4140 ttgccagcgg cttaccatcc agcgccacca tccagtgcag gagctcgtta tcgctatgac 4200 ggaacaggta ttcgctggtc acttcgatgg tttgcccgga taaacggaac tggaaaaact 4260 gctgctggtg ttttgcttcc gtcagcgctg gatgcggcgt gcggtcggca aagaccagac 4320 cgttcataca gaactggcga tcgttcggcg tatcgccaaa atcaccgccg taagccgacc 4380 acgggttgcc gttttcatca tatttaatca gcgactgatc cacccagtcc cagacgaagc 4440 cgccctgtaa acggggatac tgacgaaacg cctgccagta tttagcgaaa ccgccaagac 4500 tgttacccat cgcgtgggcg tattcgcaaa ggatcagcgg gcgcgtctct ccaggtagcg 4560 aaagccattt tttgatggac catttcggca cagccgggaa gggctggtct tcatccacgc 4620 gcgcgtacat cgggcaaata atatcggtgg ccgtggtgtc ggctccgccg ccttcatact 4680 gcaccgggcg ggaaggatcg acagatttga tccagcgata cagcgcgtcg tgattagcgc 4740 cgtggcctga ttcattcccc agcgaccaga tgatcacact cgggtgatta cgatcgcgct 4800 gcaccattcg cgttacgcgt tcgctcatcg ccggtagcca gcgcggatca tcggtcagac 4860 gattcattgg caccatgccg tgggtttcaa tattggcttc atccaccaca tacaggccgt 4920 agcggtcgca cagcgtgtac cacagcggat ggttcggata atgcgaacag cgcacggcgt 4980 taaagttgtt ctgcttcatc agcaggatat cctgcaccat cgtctgctca tccatgacct 5040 gaccatgcag aggatgatgc tcgtgacggt taacgcctcg aatcagcaac ggcttgccgt 5100 tcagcagcag cagaccattt tcaatccgca cctcgcggaa accgacatcg caggcttctg 5160 cttcaatcag cgtgccgtcg gcggtgtgca gttcaaccac cgcacgatag agattcggga 5220 tttcggcgct ccacagtttc gggttttcga cgttgagacg tagtgtgacg cgatcggcat 5280 aaccaccacg ctcatcgata atttcaccgc cgaaaggcgc ggtgccgctg gcgacctgcg 5340 tttcaccctg ccataaagaa actgttaccc gtaggtagtc acgcaactcg ccgcacatct 5400 gaacttcagc ctccagtaca gcgcggctga aatcatcatt aaagcgagtg gcaacatgga 5460 aatcgctgat ttgtgtagtc ggtttatgca gcaacgagac gtcacggaaa atgccgctca 5520 tccgccacat atcctgatct tccagataac tgccgtcact ccaacgcagc accatcaccg 5580 cgaggcggtt ttctccggcg cgtaaaaatg cgctcaggtc aaattcagac ggcaaacgac 5640 tgtcctggcc gtaaccgacc cagcgcccgt tgcaccacag atgaaacgcc gagttaacgc 5700 catcaaaaat aattcgcgtc tggccttcct gtagccagct ttcatcaaca ttaaatgtga 5760 gcgagtaaca acccgtcgga ttctccgtgg gaacaaacgg cggattgacc gtaatgggat 5820 aggttacgtt ggtgtagatg ggcgcatcgt aaccgtgcat ctgccagttt gaggggacga 5880 cgacagtatc ggcctcagga agatcgcact ccagccagct ttccggcacc gcttctggtg 5940 ccggaaacca ggcaaagcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 6000 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 6060 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg ggatccctcg 6120 aggaattcat ttatagcata gaaaaaaaca aaatgaaatt ctactatatt tttacataca 6180 tatattctaa atatgaaagt ggtgattgtg actagcgtag catcgcttct agacatatac 6240 tatatagtaa taccaatact caagactacg aaactgatac aatctcttat catgtgggta 6300 atgttctcga tgtcgaatag ccatatgccg gtagttgcga tatacataaa ctgatcacta 6360 attccaaacc cacccgcttt ttatagtaag tttttcaccc ataaataata aatacaataa 6420 ttaatttctc gtaaaagtag aaaatatatt ctaatttatt gcacggtaag gaagtagaat 6480 cataaagaac agtgacggat cccgtaaaac gacggccagt gagcgcgcgt aatacgactc 6540 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga 6600 tatcgaattc ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga 6660 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat 6720 agctgttggg aattctgtga gcgtatggca aacgaaggaa aaattagtta tagtagccgc 6780 actcgatggg acatttcaac gtaaaccgtt taataatatt ttgaatctta ttccattatc 6840 tgaaatggtg gtaaaactaa ctgctgtgtg tatgaaatgc tttaaggagg cttccttttc 6900 taaacgattg ggtgaggaaa ccgagataga aataatagga ggtaatgata tgtatcaatc 6960 ggtgtgtaga aagtgttaca tcgactcata atattatatt ttttatctaa aaaactaaaa 7020 ataaacattg attaaatttt aatataatac ttaaaaatgg atgttgtgtc gttagataaa 7080 ccgtttatgt attttgagga aattgataat gagttagatt acgaaccaga aagtgcaaat 7140 gaggtcgcaa aaaaactgcc gtatcaagga cagttaaaac tattactagg agaattattt 7200 tttcttagta agttacagcg acacggtata ttagatggtg ccaccgtagt gtatatagga 7260 tctgctcccg gtacacatat acgttatttg agagatcatt tctataattt aggagtgatc 7320 atcaaatgga tgctaattga cggccgccat catgatccta ttttaaatgg attgcgtgat 7380 gtgactctag tgactcggtt cgttgatgag gaatatctac gatccatcaa aaaacaactg 7440 catccttcta agattatttt aatttctgat gtgagatcca aacgaggagg aaatgaacct 7500 agtacggcgg atttactaag taattacgct ctacaaaatg tcatgattag tattttaaac 7560 cccgtggcgt ctagtcttaa atggagatgc ccgtttccag atcaatggat caaggacttt 7620 tatatcccac acggtaataa aatgttacaa ccttttgctc cttcatattc agggccgtcg 7680 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac 7740 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 7800 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcttt acgcatctgt 7860 gcggtatttc acaccgcata tggtgcactc tcagtaccat ctgctctgat gccgcatagt 7920 taagccagta cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc 7980 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag 8040 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg 8100 cgaggcag 8108 <210> SEQ ID NO 78 <211> LENGTH: 2091 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 78 atgtccgtgc gcgggcatgc cgtacgccgg aggcgcgcct ccacccggtc ccatgccccg 60 tccgcgcatc gcgccgactc gcccgtggag gacgagcccg agggcggtgg agtcgggtta 120 atggggtacc tgcgggcggt gtttaacgtg gacgacgaca gcgaggtcga ggccgcgggg 180 gagatggcga gcgaagagcc gcccccgcgc cgtcgccggg aggcccgcgg tcaccccggg 240 tcccgacgcg cgtccgaggc ccgggcggcg gcgccccccc gccgggcgtc ctttccgcgc 300 cccaggtccg ttacggccag gagccagtcc gttcgcggac gccgggacag cgccatcacg 360 cgggccccgc ggggaggcta cctgggcccg atggacccac gcgacgtttt ggggcgggtg 420 ggcggttcgc gggtggtgcc ctcgccgctg ttcctggacg agctcaacta cgaggaggac 480 gactaccccg ccgccgtcgc gcacgatgac ggccccgggg cgcggccttc cgcgacggtc 540 gagattctcg cgggccgcgt gtcgggcccg gagctgcagg cggcattccc cctggaccgc 600 ctgacccccc gagtcgccgc gtgggacgag tccgtgcgct cggccctggc cctgggacat 660 ccggccgggt tctacccgtg tccggatagc gcgttcgggc tgtcgcgcgt gggggtcatg 720 cactttgcct ccccggccga cccaaaggtg tttttccgcc agacgctgca gcagggcgag 780 gcgctggcct ggtacgtcac gggcgacgcg atcctcgacc tgacggatcg gcgggcaaaa 840 accagcccct cccgcgcgat gggttttctg gtggacgcca tcgtgcgggt ggcgatcaac 900 gggtgggtct gcgggacgcg cctgcacacg gaggggcgcg gctcggagct cgacgacagg 960 gcggccgagc tccgacggca gttcgcgagc ctcacggcgt tgcggcccgt gggggccgcc 1020 gccgtgccgc tgctcagcgc gggaggggcc gcgccccccc accccggccc cgacgccgcg 1080 gtctttcgca gttcgctggg gtccctgctg tactggcccg gggtgcgcgc gctcctgggg 1140 cgcgactgtc gcgtggccgc ccgctacgcg gggcgcatga cgtacatcgc caccggggct 1200 ctgctcgccc gcttcaaccc cggcgccgtc aaatgcgtgc tcccgcggga ggccgcgttt 1260 gcggggcgcg tcctggacgt gctggcggtc ctggcggagc agacggtcca gtggctctcg 1320 gtggtcgtgg gggcgcgcct gcacccgcac tccgcccacc ccgcgtttgc ggacgtggag 1380 caggaggcgc tgtttcgcgc cctgcccctg ggcagccccg gggtcgtggc ggccgagcac 1440 gaggcgctgg gcgacaccgc ggcgcgccgc ctgctcgcca ccagcgggct gaacgccgtg 1500 ctgggcgcgg ccgtgtacgc gctgcacacg gccctggcga ccgttaccct gaaatacgcc 1560 ctggcctgcg gggacgcgcg ccggcgcagg gacgacgcgg cggccgcgcg cgccgtgctg 1620 gcgacggggc tcatcctgca gcggctgctg ggcctggccg acacggtggt cgcgtgcgtg 1680 gccctggccg cgtttgacgg cgggtcgacg gcccccgagg tgggcacgta cacccccctg 1740 cgctacgcgt gcgtcctccg cgcgacccag cccctgtacg cgcggaccac ccccgccaaa 1800 ttttgggcgg acgtgcgcgc cgccgcggaa cacgtggacc ttcgccccgc gtcctcggcg 1860 ccccgggcgc ccgtgagcgg gacggcagac cccgccttcc tgctcgaaga cctggcggcc 1920 ttcccccccg cccccctgaa tagcgagtcc gtgctggggc cgcgggtccg cgtcgtggac 1980 atcatggcgc agtttcggaa actgctcatg ggcgacgagg agaccgccgc cctccgggcg 2040 cacgtgtccg ggaggcgcgc gaccgggctg ggcggcccgc cacgcccata g 2091 <210> SEQ ID NO 79 <211> LENGTH: 1110 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 79 atgagtcagt gggggcccag ggcgatcctt gtccagacgg acagcaccaa ccggaatgcc 60 gatggggact ggcaagcggc cgtagctatt cgcgggggcg gagtcgttca actgaacatg 120 gtcaacaaac gcgccgtgga ttttaccccg gcagaatgcg gggactccga atgggccgtg 180 ggccgcgtct ctctgggcct gcgaatggca atgccgcggg acttctgcgc gattattcac 240 gcccccgcgg tatccggccc cgggccccac gtgatgctcg gtctcgtcga ctcgggctac 300 cgcggaaccg tcctggccgt ggtcgtagcc ccgaacggga cgcgcgggtt tgcccccggg 360 gccctccggg tcgacgtgac gtttctggac atccgggcca cccccccgac cctcaccgag 420 ccgagctccc tgcaccggtt tccgcagttg gcgccgtccc cgctggcagg gttacgagaa 480 gatccttggt tggacggggc gctcgcgacc gccggggggg cggtggccct gccggccaga 540 cggcgcgggg gatcgctggt ctacgcgggc gagctaacgc aggtgaccac cgagcacggc 600 gactgcgtgc acgaggcgcc cgcctttctg ccaaagcgcg aggaggacgc aggctttgac 660 attctcatcc accgagccgt gaccgtcccg gccaacggcg ccacggtcat acagccgtcc 720 ctccgcgtat tgcgcgcggc cgacggacca gaggcctgct atgtgctggg gcggtcgtcg 780 ctcaatgcca ggggcctcct ggtcatgcct acgcgctggc cctccgggca cgcctgtgcg 840 tttgttgtat gtaacctgac cggagtcccg gtgaccctac aagccgggtc caaggtcgcc 900 cagctgctcg tcgcggggac ccacgccctc ccctggatcc cccccgacaa catccacgag 960 gacggcgcat tccgggccta ccccagaggg gttccggacg cgaccgccac cccccgagac 1020 ccgccgattt tggtgtttac gaacgagttt gacgcggacg cccccccaag caagcggggg 1080 gccggggggt ttggctccac tggcatctag 1110 <210> SEQ ID NO 80 <211> LENGTH: 228 <212> TYPE: DNA <213> ORGANISM: Homo sapiens-ubiquitin UL49-HSV-2 <400> SEQUENCE: 80 atgcagatct tcgtgaagac tctgactggt aagaccatca ccctcgaggt ggagcccagt 60 gacaccatcg agaatgtcaa ggcaaagatc caagataagg aaggcattcc tcctgatcag 120 cagaggttga tctttgccgg aaaacagctg gaagatggtc gtaccctgtc tgactacaac 180 atccagaaag agtccacctt gcacctggta ctccgtctca gaggtggg 228 <210> SEQ ID NO 81 <211> LENGTH: 903 <212> TYPE: DNA <213> ORGANISM: Homo sapiens-ubiquitin UL49-HSV-2 <400> SEQUENCE: 81 atgacctctc gccgctccgt caagtcgtgt ccgcgggaag cgccgcgcgg gacccacgag 60 gagctgtact atggcccggt ctccccggcg gacccagaga gtccgcgcga cgacttccgc 120 cgcggcgctg gcccgatgcg cgcgcgcccg aggggcgagg ttcgctttct ccattatgac 180 gaggctgggt atgccctcta ccgggactcg tcttcggacg acgacgagtc ccgggatacc 240 gcgcgaccgc gtcgttcggc gtccgtcgcg ggctctcacg gccccggccc cgcgcgcgct 300 cctccacccc ccgggggccc cgtgggcgcc ggcgggcgct cgcacgcccc tcccgcgcgg 360 acccccaaaa tgacgcgcgg ggcgcctaag gcctccgcga ccccggcgac cgaccccgcc 420 cgcggcaggc gacccgccca ggccgactcc gccgtgctcc tagacgcccc cgctcccacg 480 gcctcgggaa gaaccaagac acccgcccag ggactggcca agaagctgca cttcagcacc 540 gccccaccga gccccacggc gccgtggacc ccccgggtgg ccgggttcaa caagcgcgtc 600 ttctgcgccg cggtcgggcg cctggcggcc acgcacgccc ggctggcggc ggtacagctg 660 tgggacatgt cgcggccgca caccgacgaa gacctcaacg agctcctcga cctcaccacc 720 attcgcgtga cggtctgcga gggcaagaac ctcctgcagc gcgcgaacga gttggtgaat 780 cccgacgcgg cgcaggacgt cgacgcgacc gcggccgccc ggggccgccc cgcggggcgt 840 gccgccgcga ccgcacgggc ccccgcccgc tcggcttccc gtccccgccg ccccctcgag 900 tag 903 <210> SEQ ID NO 82 <211> LENGTH: 1113 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 82 atgagtcagt gggggcccag ggcgatcctt gtccagacgg acagcaccaa ccggaatgcc 60 gatggggact ggcaagcggc cgtagctatt cgcgggggcg gagtcgttca actgaacatg 120 gtcaacaaac gcgccgtgga ttttaccccg gcagaatgcg gggactccga atgggccgtg 180 ggccgcgtct ctctgggcct gcgaatggca atgccgcggg acttctgcgc gattattcac 240 gcccccgcgg tatccggccc cgggccccac gtgatgctcg gtctcgtcga ctcgggctac 300 cgcggaaccg tcctggccgt ggtcgtagcc ccgaacggga cgcgcgggtt tgcccccggg 360 gccctccggg tcgacgtgac gtttctggac atccgggcca cccccccgac cctcaccgag 420 ccgagctccc tgcaccggtt tccgcagttg gcgccgtccc cgctggcagg gttacgagaa 480 gatccttggt tggacggggc gctcgcgacc gccggggggg cggtggccct gccggccaga 540 cggcgcgggg gatcgctggt ctacgcgggc gagctaacgc aggtgaccac cgagcacggc 600 gactgcgtgc acgaggcgcc cgcctttctg ccaaagcgcg aggaggacgc aggctttgac 660 attctcatcc accgagccgt gaccgtcccg gccaacggcg ccacggtcat acagccgtcc 720 ctccgcgtat tgcgcgcggc cgacggacca gaggcctgct atgtgctggg gcggtcgtcg 780 ctcaatgcca ggggcctcct ggtcatgcct acgcgctggc cctccgggca cgcctgtgcg 840 tttgttgtat gtaacctgac cggagtcccg gtgaccctac aagccgggtc caaggtcgcc 900 cagctgctcg tcgcggggac ccacgccctc ccctggatcc cccccgacaa catccacgag 960 gacggcgcat tccgggccta ccccagaggg gttccggacg cgaccgccac cccccgagac 1020 ccgccgattt tggtgtttac gaacgagttt gacgcggacg cccccccaag caagcggggg 1080 gccggggggt ttggctccac tggcatctag tga 1113 <210> SEQ ID NO 83 <211> LENGTH: 927 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 83 atgcagcatc accaccatca ccaccacacc tctcgccgct ccgtcaagtc gtgtccgcgg 60 gaagcgccgc gcgggaccca cgaggagctg tactatggcc cggtctcccc ggcggaccca 120 gagagtccgc gcgacgactt ccgccgcggc gctggcccga tgcgcgcgcg cccgaggggc 180 gaggttcgct ttctccatta tgacgaggct gggtatgccc tctaccggga ctcgtcttcg 240 gacgacgacg agtcccggga taccgcgcga ccgcgtcgtt cggcgtccgt cgcgggctct 300 cacggccccg gccccgcgcg cgctcctcca ccccccgggg gccccgtggg cgccggcggg 360 cgctcgcacg cccctcccgc gcggaccccc aaaatgacgc gcggggcgcc taaggccccc 420 gcgaccccgg cgaccgaccc cgcccgcggc aggcgacccg cccaggccga ctccgccgtg 480 ctcctagacg cccccgctcc cacggcctcg ggaagaacca agacacccgc ccagggactg 540 gccaagaagc tgcacttcag caccgcccca ccgagcccca cggcgccgtg gaccccccgg 600 gtggccgggt tcaacaagcg cgtcttctgc gccgcggtcg ggcgcctggc ggccacgcac 660 gcccggctgg cggcggtaca gctgtgggac atgtcgcggc cgcacaccga cgaagacctc 720 aacgagctcc tcgacctcac caccattcgc gtgacggtct gcgagggcaa gaacctcctg 780 cagcgcgcga acgagttggt gaatcccgac gcggcgcagg acgtcgacgc gaccgcggcc 840 gcccggggcc gccccgcggg gcgtgccgcc gcgaccgcac gggcccccgc ccgctcggct 900 tcccgtcccc gccgccccct cgagtag 927 <210> SEQ ID NO 84 <211> LENGTH: 4149 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 84 atgcagcatc accaccatca ccacgccgct cctgcccgcg accccccggg ttaccggtac 60 gccgcggcca tggtgcccac cggctccatc ctgagtacga tcgaggtggc gtcccaccgc 120 agactctttg attttttcgc ccgcgtgcgc tccgacgaaa acagcctgta tgacgtagag 180 tttgacgccc tgctggggtc ctactgcaac accctgtcgc tcgtgcgctt tctggagctc 240 ggcctgtccg tggcgtgcgt gtgcaccaag ttcccggagc tggcttacat gaacgaaggg 300 cgtgtgcagt tcgaggtcca ccagcccctc atcgcccgcg acggcccgca ccccgtcgag 360 cagcccgtgc ataattacat gacgaaggtc atcgaccgcc gggccctgaa cgccgccttc 420 agcctggcca ccgaggccat tgccctgctc acgggggagg ccctggacgg gacgggcatt 480 agcctgcatc gccagctgcg cgccatccag cagctcgcgc gcaacgtcca ggccgtcctg 540 ggggcgtttg agcgcggcac ggccgaccag atgctgcacg tgctgttgga gaaggcgcct 600 cccctggccc tgctgttgcc catgcaacga tatctcgaca acgggcgcct ggcgaccagg 660 gtcgcccggg cgaccctggt cgccgagctg aagcggagct tttgcgacac gagcttcttc 720 ctgggcaagg cgggccatcg ccgcgaggcc atcgaggcct ggctcgtgga cctgaccacg 780 gcgacgcagc cgtccgtggc cgtgccccgc ctgacgcacg ccgacacgcg cgggcggccg 840 gtcgacgggg tgctggtcac caccgccgcc atcaaacagc gcctcctgca gtccttcctg 900 aaggtggagg acaccgaggc cgacgtgccg gtgacctacg gcgagatggt cttgaacggg 960 gccaacctcg tcacggcgct ggtgatgggc aaggccgtgc ggagcctgga cgacgtgggc 1020 cgccacctgc tggagatgca ggaggagcaa ctcgaggcga accgggagac gctggatgaa 1080 ctcgaaagcg ccccccagac aacgcgcgtg cgcgcggatc tggtggccat aggcgacagg 1140 ctggtcttcc tggaggccct ggagaagcgc atctacgccg ccaccaacgt gccctacccc 1200 ctggtgggcg ccatggacct gacgttcgtc ctgcccctgg ggctgttcaa cccggccatg 1260 gagcgcttcg ccgcgcacgc cggggacctg gtgcccgccc ccggccaccc ggagccccgc 1320 gcgttccctc cccggcagct gtttttttgg ggaaaggacc accaggttct gcggctgtcc 1380 atggagaacg cggtcgggac cgtgtgtcat ccttcgctca tgaacatcga cgcggccgtc 1440 gggggcgtga accacgcccc cgtcgaggcc gcgaacccgt acggggcgta cgtcgcggcc 1500 ccggccggcc ccggcgcgga catgcagcag cgttttctga acgcctggcg gcagcgcctc 1560 gcccacggcc gggtccggtg ggtcgccgag tgccagatga ccgcggagca gttcatgcag 1620 cccgacaacg ccaacctggc tctggagctg caccccgcgt tcgacttctt cgcgggcgtg 1680 gccgacgtcg agcttcccgg cggcgaagtc cccccggccg gtccgggggc gatccaggcc 1740 acctggcgcg tggtcaacgg caacctgccc ctggcgctgt gtccggtggc gtttcgtgac 1800 gcccggggcc tggagctcgg cgttggccgc cacgccatgg cgccggctac catagccgcc 1860 gtccgcgggg cgttcgagga ccgcagctac ccggcggtgt tttacctgct gcaagccgcg 1920 attcacggca gcgagcacgt gttctgcgcc ctggcgcggc tcgtgactca gtgcatcacc 1980 agctactgga acaacacgcg atgcgcggcg ttcgtgaacg actactcgct ggtctcgtac 2040 atcgtgacct acctcggggg cgacctcccc gaggagtgca tggccgtgta tcgggacctg 2100 gtggcccacg tcgaggccct ggcccagctg gtggacgact ttaccctgcc gggcccggag 2160 ctgggcgggc aggctcaggc cgagctgaat cacctgatgc gcgacccggc gctgctgccg 2220 cccctcgtgt gggactgcga cggccttatg cgacacgcgg ccctggaccg ccaccgagac 2280 tgccggattg acgcgggggg gcacgagccc gtctacgcgg cggcgtgcaa cgtggcgacg 2340 gccgacttta accgcaacga cggccggctg ctgcacaaca cccaggcccg cgcggtcgac 2400 gccgccgacg accggccgca ccggccggcc gactggaccg tccaccacaa aatctactat 2460 tacgtgctgg tgccggcctt ctcgcggggg cgctgctgca ccgcgggggt ccgcttcgac 2520 cgcgtgtacg ccacgctgca gaacatggtg gtcccggaga tcgcccccgg tgaggagtgc 2580 ccgagcgatc ccgtgaccga ccccgcccac ccgctgcatc ccgccaatct ggtggccaac 2640 acggtcaacg ccatgttcca caacgggcgc gtcgtcgtcg acgggcccgc catgctcacg 2700 ctgcaggtgc tggcgcacaa catggccgag cgcacgacgg cgctgctgtg ctccgcggcg 2760 cccgacgcgg gcgccaacac cgcgtcgacg gccaacatgc gcatcttcga cggggcgctg 2820 cacgccggcg tgctgctcat ggccccccag cacctggacc acaccatcca aaatggcgaa 2880 tacttctacg tcctgcccgt ccacgcgctg tttgcgggcg ccgaccacgt ggccaacgcg 2940 cccaacttcc ccccggccct gcgcgacctg gcgcgccacg tccccctggt ccccccggcc 3000 ctgggggcca actacttctc ctccatccgc cagcccgtgg tgcagcacgc ccgcgagagc 3060 gcggcggggg agaacgcgct gacctacgcg ctcatggcgg ggtacttcaa gatgagcccc 3120 gtggccctgt atcaccagct caagacgggc ctccaccccg ggttcgggtt caccgtcgtg 3180 cggcaggacc gcttcgtgac cgagaacgtg ctgttttccg agcgcgcgtc ggaggcgtac 3240 tttctgggcc agctccaggt ggcccgccac gaaacgggcg ggggggtcag cttcacgctc 3300 acccagccgc gcggaaacgt ggacctgggt gtgggctaca ccgccgtcgc ggccacggcc 3360 accgtccgca accccgttac ggacatgggc aacctccccc aaaactttta cctcggccgc 3420 ggggcccccc cgctgctaga caacgcggcc gccgtgtacc tgcgcaacgc ggtcgtggcg 3480 ggaaaccggc tggggccggc ccagcccctc ccggtctttg gctgcgccca ggtgccgcgg 3540 cgcgccggca tggaccacgg gcaggatgcc gtgtgtgagt tcatcgccac ccccgtggcc 3600 acggacatca actactttcg ccggccctgc aacccgcggg gacgcgcggc cggcggcgtg 3660 tacgcggggg acaaggaggg ggacgtcata gccctcatgt acgaccacgg ccagagcgac 3720 ccggcgcggc ccttcgcggc cacggccaac ccgtgggcgt cgcagcggtt ctcgtacggg 3780 gacctgctgt acaacggggc ctatcacctc aacggggcct cgcccgtcct cagcccctgc 3840 ttcaagttct tcaccgcggc cgacatcacg gccaaacatc gctgcctgga gcgtcttatc 3900 gtggaaacgg gatcggcggt atccacggcc accgctgcca gcgacgtgca gtttaagcgc 3960 ccgccggggt gccgcgagct cgtggaagac ccgtgcggcc tgtttcagga agcctacccg 4020 atcacctgcg ccagcgaccc cgccctgcta cgcagcgccc gcgatgggga ggcccacgcg 4080 cgagagaccc actttacgca gtatctcatc tacgacgcct ccccgctaaa gggcctgtct 4140 ctgtaatga 4149 <210> SEQ ID NO 85 <211> LENGTH: 1623 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 85 atgcagcatc accaccatca ccacgagctc agctatgcca ccaccctgca ccaccgggac 60 gttgtgtttt acgtcacggc agacagaaac cgcgcctact ttgtgtgcgg ggggtccgtt 120 tattccgtag ggcggcctcg ggattctcag ccgggggaaa ttgccaagtt tggcctggtg 180 gtccggggga caggccccaa agaccgcatg gtcgccaact acgtacgaag cgagctccgc 240 cagcgcggcc tgcgggacgt gcggcccgtg ggggaggacg aggtgttcct ggacagcgtg 300 tgtctgctaa acccgaacgt gagctccgag cgagacgtga ttaataccaa cgacgttgaa 360 gtgctggacg aatgcctggc cgaatactgc acctcgctgc gaaccagccc gggggtgctg 420 gtgaccgggg tgcgcgtgcg cgcgcgagac agggtcatcg agctatttga gcacccggcg 480 atcgtcaaca tttcctcgcg cttcgcgtac accccctccc cctacgtatt cgccctggcc 540 caggcgcacc tcccccggct cccgagctcg ctggagcccc tggtgagcgg cctgtttgac 600 ggcattcccg ccccgcgcca gcccctggac gcccgcgacc ggcgcacgga tgtcgtgatc 660 acgggcaccc gcgcccccag accgatggcc gggaccgggg ccgggggcgc gggggccaag 720 cgggccaccg tcagcgagtt cgtgcaagtg aagcacatcg accgtgttgt gtccccgagc 780 gtctcttccg cccccccgcc gagcgccccc gacgcgagtc tgccgccccc ggggctccag 840 gaggccgccc cgccgggccc cccgctcagg gagctgtggt gggtgttcta cgccggcgac 900 cgggcgctgg aggagcccca cgccgagtcg ggattgacgc gcgaggaggt ccgcgccgtg 960 catgggttcc gggagcaggc gtggaagctg tttgggtcgg tgggggctcc gcgggcgttt 1020 ctcggggccg cgctggccct gagcccgacc caaaagctcg ccgtctacta ctatctcatc 1080 caccgggagc ggcgcatgtc ccccttcccc gcgctcgtgc ggctcgtcgg tcggtacatc 1140 cagcgccacg gcctgtacgt tcccgcgccc gacgaaccga cgttggccga tgccatgaac 1200 gggctgttcc gcgacgcgct ggcggccggg accgtggccg agcagctcct catgttcgac 1260 ctcctcccgc ccaaggacgt gccggtgggg agcgacgcgc gggccgacag cgccgccctg 1320 ctgcgctttg tggactcgca acgcctgacc ccgggggggt ccgtctcgcc cgagcacgtc 1380 atgtacctcg gcgcgttcct gggcgtgttg tacgccggcc acggacgcct ggccgcggcc 1440 acgcataccg cgcgcctgac gggcgtgacg tccctggtcc tgaccgtggg ggacgtcgac 1500 cggatgtccg cgtttgaccg cgggccggcg ggggcggctg gccgcacgcg aaccgccggg 1560 tacctggacg cgctgcttac cgtttgcctg gctcgcgccc agcacggcca gtctgtgtga 1620 tga 1623 <210> SEQ ID NO 86 <211> LENGTH: 2211 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 86 atgcagcatc accaccatca ccacgtgtct atcgaaggtc gtgctagctc tggtggcagc 60 ggtctggttc cgcgtggtag ctctggttcg ggggacgacg acgacaaatc tagtaggcac 120 tccgtgcgcg ggcatgccgt acgccggagg cgcgcctcca cccggtccca tgccccgtcc 180 gcgcatcgcg ccgactcgcc cgtggaggac gagcccgagg gcggtggagt cgggttaatg 240 gggtacctgc gggcggtgtt taacgtggac gacgacagcg aggtcgaggc cgcgggggag 300 atggcgagcg aagagccgcc cccgcgccgt cgccgggagg cccgcggtca ccccgggtcc 360 cgacgcgcgt ccgaggcccg ggcggcggcg cccccccgcc gggcgtcctt tccgcgcccc 420 aggtccgtta cggccaggag ccagtccgtt cgcggacgcc gggacagcgc catcacgcgg 480 gccccgcggg gaggctacct gggcccgatg gacccacgcg acgttttggg gcgggtgggc 540 ggttcgcggg tggtgccctc gccgctgttc ctggacgagc tcagctacga ggaggacgac 600 taccccgccg ccgtcgcgca cgatgacggc gccggggcgc ggcctcccgc gacggtcgag 660 attctcgcgg gccgcgtgtc gggcccggag ctgcaggcgg cattccccct ggaccgcctg 720 accccccgag tcgccgcgtg ggacgagtcc gtgcgctcgg ccctggccct gggacatccg 780 gccgggttct acccgtgtcc ggatagcgcg ttcgggctgt cgcgcgtggg ggtcatgcac 840 tttgcctccc cggccgaccc aaaggtgttt ttccgccaga cgctgcagca gggcgaggcg 900 ctggcctggt acgtcacggg cgacgcgatc ctcgacctga cggatcggcg ggcaaaaacc 960 agcccctccc gcgcgatggg ttttctggtg gacgccatcg tgcgggtggc gatcaacggg 1020 tgggtctgcg ggacgcgcct gcacacggag gggcgcggct cggagctcga cgacagggcg 1080 gccgagctcc gacggcagtt cgcgagcctc acggcgttgc ggcccgtggg ggccgccgcc 1140 gtgccgctgc tcagcgcggg aggggccgcg cccccccacc ccggccccga cgccgcggtc 1200 tttcgcagtt cgctggggtc cctgctgtac tggcccgggg tgcgcgcgct cctggggcgc 1260 gactgtcgcg tggccgcccg ctacgcgggg cgcatgacgt acatcgccac cggggctctg 1320 ctcgcccgct tcaaccccgg cgccgtcaaa tgcgtgctcc cgcgggaggc cgcgtttgcg 1380 gggcgcgtcc tggacgtgct ggcggtcctg gcggagcaga cggtccagtg gctctcggtg 1440 gtcgtggggg cgcgcctgca cccgcactcc gcccaccccg cgtttgcgga cgtggagcag 1500 gaggcgctgt ttcgcgccct gcccctgggc agccccgggg tcgtggcggc cgagcacgag 1560 gcgctgggcg acaccgcggc gcgccgcctg ctcgccacca gcgggctgaa cgccgtgctg 1620 ggcgcggccg tgtacgcgct gcacacggcc ctggcgaccg ttaccctgaa atacgccctg 1680 gcctgcgggg acgcgcgccg gcgcagggac gacgcggcgg ccgcgcgcgc cgtgctggcg 1740 acggggctca tcctgcagcg gctgctgggc ctggccgaca cggtggtcgc gtgcgtggcc 1800 ctggccgcgt ttgacggcgg gtcgacggcc cccgaggtgg gcacgtacac ccccctgcgc 1860 tacgcgtgcg tcctccgcgc gacccagccc ctgtacgcgc ggaccacccc cgccaaattt 1920 tgggcggacg tgcgcgccgc cgcggaacac gtggaccttc gccccgcgtc ctcggcgccc 1980 cgggcgcccg tgagcgggac ggcagacccc gccttcctgc tcgaagacct ggcggccttc 2040 ccccccgccc ccctgaatag cgagtccgtg ctggggccgc gggtccgcgt cgtggacatc 2100 atggcgcagt ttcggaaact gctcatgggc gacgaggaga ccgccgccct ccgggcgcac 2160 gtgtccggga ggcgcgcgac cgggctgggc ggcccgccac gcccatagtg a 2211 <210> SEQ ID NO 87 <211> LENGTH: 2118 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 87 atgcagcatc accaccatca ccaccactcc gtgcgcgggc atgccgtacg ccggaggcgc 60 gcctccaccc ggtcccatgc cccgtccgcg catcgcgccg actcgcccgt ggaggacgag 120 cccgagggcg gtggagtcgg gttaatgggg tacctgcggg cggtgtttaa cgtggacgac 180 gacagcgagg tcgaggccgc gggggagatg gcgagcgaag agccgccccc gcgccgtcgc 240 cgggaggccc gcggtcaccc cgggtcccga cgcgcgtccg aggcccgggc ggcggcgccc 300 ccccgccggg cgtcctttcc gcgccccagg tccgttacgg ccaggagcca gtccgttcgc 360 ggacgccggg acagcgccat cacgcgggcc ccgcggggag gctacctggg cccgatggac 420 ccacgcgacg ttttggggcg ggtgggcggt tcgcgggtgg tgccctcgcc gctgttcctg 480 gacgagctca gctacgagga ggacgactac cccgccgccg tcgcgcacga tgacggcgcc 540 ggggcgcggc ctcccgcgac ggtcgagatt ctcgcgggcc gcgtgtcggg cccggagctg 600 caggcggcat tccccctgga ccgcctgacc ccccgagtcg ccgcgtggga cgagtccgtg 660 cgctcggccc tggccctggg acatccggcc gggttctacc cgtgtccgga tagcgcgttc 720 gggctgtcgc gcgtgggggt catgcacttt gcctccccgg ccgacccaaa ggtgtttttc 780 cgccagacgc tgcagcaggg cgaggcgctg gcctggtacg tcacgggcga cgcgatcctc 840 gacctgacgg atcggcgggc aaaaaccagc ccctcccgcg cgatgggttt tctggtggac 900 gccatcgtgc gggtggcgat caacgggtgg gtctgcggga cgcgcctgca cacggagggg 960 cgcggctcgg agctcgacga cagggcggcc gagctccgac ggcagttcgc gagcctcacg 1020 gcgttgcggc ccgtgggggc cgccgccgtg ccgctgctca gcgcgggagg ggccgcgccc 1080 ccccaccccg gccccgacgc cgcggtcttt cgcagttcgc tggggtccct gctgtactgg 1140 cccggggtgc gcgcgctcct ggggcgcgac tgtcgcgtgg ccgcccgcta cgcggggcgc 1200 atgacgtaca tcgccaccgg ggctctgctc gcccgcttca accccggcgc cgtcaaatgc 1260 gtgctcccgc gggaggccgc gtttgcgggg cgcgtcctgg acgtgctggc ggtcctggcg 1320 gagcagacgg tccagtggct ctcggtggtc gtgggggcgc gcctgcaccc gcactccgcc 1380 caccccgcgt ttgcggacgt ggagcaggag gcgctgtttc gcgccctgcc cctgggcagc 1440 cccggggtcg tggcggccga gcacgaggcg ctgggcgaca ccgcggcgcg ccgcctgctc 1500 gccaccagcg ggctgaacgc cgtgctgggc gcggccgtgt acgcgctgca cacggccctg 1560 gcgaccgtta ccctgaaata cgccctggcc tgcggggacg cgcgccggcg cagggacgac 1620 gcggcggccg cgcgcgccgt gctggcgacg gggctcatcc tgcagcggct gctgggcctg 1680 gccgacacgg tggtcgcgtg cgtggccctg gccgcgtttg acggcgggtc gacggccccc 1740 gaggtgggca cgtacacccc cctgcgctac gcgtgcgtcc tccgcgcgac ccagcccctg 1800 tacgcgcgga ccacccccgc caaattttgg gcggacgtgc gcgccgccgc ggaacacgtg 1860 gaccttcgcc ccgcgtcctc ggcgccccgg gcgcccgtga gcgggacggc agaccccgcc 1920 ttcctgctcg aagacctggc ggccttcccc cccgcccccc tgaatagcga gtccgtgctg 1980 gggccgcggg tccgcgtcgt ggacatcatg gcgcagtttc ggaaactgct catgggcgac 2040 gaggagaccg ccgccctccg ggcgcacgtg tccgggaggc gcgcgaccgg gctgggcggc 2100 ccgccacgcc catagtga 2118 <210> SEQ ID NO 88 <211> LENGTH: 939 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 88 atgcagcatc accaccatca ccaccactcc gtgcgcgggc atgccgtacg ccggaggcgc 60 gcctccaccc ggtcccatgc cccgtccgcg catcgcgccg actcgcccgt ggaggacgag 120 cccgagggcg gtggagtcgg gttaatgggg tacctgcggg cggtgtttaa cgtggacgac 180 gacagtgagg tcgaggccgc gggggagatg gcgagcgaag agccgccccc gcgccgtcgc 240 cgggaggccc gcggtcaccc cgggtcccga cgcgcgtccg aggcccgggc ggcggcgccc 300 ccccgccggg cgtcctttcc gcgccccagg tccgttacgg ccaggagcca gtccgttcgc 360 ggacgccggg acagcgccat cacgcgggcc ccgcggggag gctacctggg cccgatggac 420 ccacgcgacg ttttggggcg ggtgggcggt tcgcgggtgg tgccctcgcc gctgttcctg 480 gacgagctca gctacgagga ggacgactac cccgccgccg tcgcgcacga tgacggcgcc 540 ggggcgcggc ctcccgcgac ggtcgagatt ctcgcgggcc gcgtgtcggg cccggagctg 600 caggcggcat tccccctgga ccgcctgacc ccccgagtcg ccgcgtggga cgagtccgtg 660 cgctcggccc tggccctggg acatccggcc gggttctacc cgtgtccgga tagcgcgttc 720 gggctgtcgc gcgtgggggt catgcacttt gcctccccgg ccgacccaaa ggtgtttttc 780 cgccagacgc tgcagcaggg cgaggcgctg gcctggtacg tcacgggcga cgcgatcctc 840 gacctgacgg atcggcgggc aaaaaccagc ccctcccgcg cgatgggctt tctggtggac 900 gccatcgtgc gggtggcgat caacgggtgg gtctgatga 939 <210> SEQ ID NO 89 <211> LENGTH: 843 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 89 atgcagcatc accaccatca ccaccacgcc gccgcacccc aggcggacgt ggcgccggtt 60 ctggacagcc agcccactgt gggaacggac cccggctacc cagtccccct agaactcacg 120 cccgagaacg cggaggcggt ggcgcggttt ctgggggacg ccgtcgaccg cgagcccgcg 180 ctcatgctgg agtacttctg tcggtgcgcc cgcgaggaga gcaagcgcgt gcccccacga 240 accttcggca gcgccccccg cctcacggag gacgactttg ggctcctgaa ctacgcgctc 300 gctgagatgc gacgcctgtg cctggacctt cccccggtcc cccccaacgc atacacgccc 360 tatcatctga gggagtatgc gacgcggctg gttaacgggt tcaaacccct ggtgcggcgg 420 tccgcccgcc tgtatcgcat cctggggatt ctggttcacc tgcgcatccg tacccgggag 480 gcctcctttg aggaatggat gcgctccaag gaggtggacc tggacttcgg gctgacggaa 540 aggcttcgcg aacacgaggc ccagctaatg atcctggccc aggccctgaa cccctacgac 600 tgtctgatcc acagcacccc gaacacgctc gtcgagcggg ggctgcagtc ggcgctgaag 660 tacgaagagt tttacctcaa gcgcttcggc gggcactaca tggagtccgt cttccagatg 720 tacacccgca tcgccgggtt cctggcgtgc cgggcgaccc gcggcatgcg ccacatcgcc 780 ctggggcgac aggggtcgtg gtgggaaatg ttcaagttct ttttccaccg cctctactaa 840 tga 843 <210> SEQ ID NO 90 <211> LENGTH: 279 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 90 Met Gln His His His His His His His Ala Ala Ala Pro Gln Ala Asp 5 10 15 Val Ala Pro Val Leu Asp Ser Gln Pro Thr Val Gly Thr Asp Pro Gly 20 25 30 Tyr Pro Val Pro Leu Glu Leu Thr Pro Glu Asn Ala Glu Ala Val Ala 35 40 45 Arg Phe Leu Gly Asp Ala Val Asp Arg Glu Pro Ala Leu Met Leu Glu 50 55 60 Tyr Phe Cys Arg Cys Ala Arg Glu Glu Ser Lys Arg Val Pro Pro Arg 65 70 75 80 Thr Phe Gly Ser Ala Pro Arg Leu Thr Glu Asp Asp Phe Gly Leu Leu 85 90 95 Asn Tyr Ala Leu Ala Glu Met Arg Arg Leu Cys Leu Asp Leu Pro Pro 100 105 110 Val Pro Pro Asn Ala Tyr Thr Pro Tyr His Leu Arg Glu Tyr Ala Thr 115 120 125 Arg Leu Val Asn Gly Phe Lys Pro Leu Val Arg Arg Ser Ala Arg Leu 130 135 140 Tyr Arg Ile Leu Gly Ile Leu Val His Leu Arg Ile Arg Thr Arg Glu 145 150 155 160 Ala Ser Phe Glu Glu Trp Met Arg Ser Lys Glu Val Asp Leu Asp Phe 165 170 175 Gly Leu Thr Glu Arg Leu Arg Glu His Glu Ala Gln Leu Met Ile Leu 180 185 190 Ala Gln Ala Leu Asn Pro Tyr Asp Cys Leu Ile His Ser Thr Pro Asn 195 200 205 Thr Leu Val Glu Arg Gly Leu Gln Ser Ala Leu Lys Tyr Glu Glu Phe 210 215 220 Tyr Leu Lys Arg Phe Gly Gly His Tyr Met Glu Ser Val Phe Gln Met 225 230 235 240 Tyr Thr Arg Ile Ala Gly Phe Leu Ala Cys Arg Ala Thr Arg Gly Met 245 250 255 Arg His Ile Ala Leu Gly Arg Gln Gly Ser Trp Trp Glu Met Phe Lys 260 265 270 Phe Phe Phe His Arg Leu Tyr 275 <210> SEQ ID NO 91 <211> LENGTH: 539 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 91 Met Gln His His His His His His Glu Leu Ser Tyr Ala Thr Thr Leu 5 10 15 His His Arg Asp Val Val Phe Tyr Val Thr Ala Asp Arg Asn Arg Ala 20 25 30 Tyr Phe Val Cys Gly Gly Ser Val Tyr Ser Val Gly Arg Pro Arg Asp 35 40 45 Ser Gln Pro Gly Glu Ile Ala Lys Phe Gly Leu Val Val Arg Gly Thr 50 55 60 Gly Pro Lys Asp Arg Met Val Ala Asn Tyr Val Arg Ser Glu Leu Arg 65 70 75 80 Gln Arg Gly Leu Arg Asp Val Arg Pro Val Gly Glu Asp Glu Val Phe 85 90 95 Leu Asp Ser Val Cys Leu Leu Asn Pro Asn Val Ser Ser Glu Arg Asp 100 105 110 Val Ile Asn Thr Asn Asp Val Glu Val Leu Asp Glu Cys Leu Ala Glu 115 120 125 Tyr Cys Thr Ser Leu Arg Thr Ser Pro Gly Val Leu Val Thr Gly Val 130 135 140 Arg Val Arg Ala Arg Asp Arg Val Ile Glu Leu Phe Glu His Pro Ala 145 150 155 160 Ile Val Asn Ile Ser Ser Arg Phe Ala Tyr Thr Pro Ser Pro Tyr Val 165 170 175 Phe Ala Leu Ala Gln Ala His Leu Pro Arg Leu Pro Ser Ser Leu Glu 180 185 190 Pro Leu Val Ser Gly Leu Phe Asp Gly Ile Pro Ala Pro Arg Gln Pro 195 200 205 Leu Asp Ala Arg Asp Arg Arg Thr Asp Val Val Ile Thr Gly Thr Arg 210 215 220 Ala Pro Arg Pro Met Ala Gly Thr Gly Ala Gly Gly Ala Gly Ala Lys 225 230 235 240 Arg Ala Thr Val Ser Glu Phe Val Gln Val Lys His Ile Asp Arg Val 245 250 255 Val Ser Pro Ser Val Ser Ser Ala Pro Pro Pro Ser Ala Pro Asp Ala 260 265 270 Ser Leu Pro Pro Pro Gly Leu Gln Glu Ala Ala Pro Pro Gly Pro Pro 275 280 285 Leu Arg Glu Leu Trp Trp Val Phe Tyr Ala Gly Asp Arg Ala Leu Glu 290 295 300 Glu Pro His Ala Glu Ser Gly Leu Thr Arg Glu Glu Val Arg Ala Val 305 310 315 320 His Gly Phe Arg Glu Gln Ala Trp Lys Leu Phe Gly Ser Val Gly Ala 325 330 335 Pro Arg Ala Phe Leu Gly Ala Ala Leu Ala Leu Ser Pro Thr Gln Lys 340 345 350 Leu Ala Val Tyr Tyr Tyr Leu Ile His Arg Glu Arg Arg Met Ser Pro 355 360 365 Phe Pro Ala Leu Val Arg Leu Val Gly Arg Tyr Ile Gln Arg His Gly 370 375 380 Leu Tyr Val Pro Ala Pro Asp Glu Pro Thr Leu Ala Asp Ala Met Asn 385 390 395 400 Gly Leu Phe Arg Asp Ala Leu Ala Ala Gly Thr Val Ala Glu Gln Leu 405 410 415 Leu Met Phe Asp Leu Leu Pro Pro Lys Asp Val Pro Val Gly Ser Asp 420 425 430 Ala Arg Ala Asp Ser Ala Ala Leu Leu Arg Phe Val Asp Ser Gln Arg 435 440 445 Leu Thr Pro Gly Gly Ser Val Ser Pro Glu His Val Met Tyr Leu Gly 450 455 460 Ala Phe Leu Gly Val Leu Tyr Ala Gly His Gly Arg Leu Ala Ala Ala 465 470 475 480 Thr His Thr Ala Arg Leu Thr Gly Val Thr Ser Leu Val Leu Thr Val 485 490 495 Gly Asp Val Asp Arg Met Ser Ala Phe Asp Arg Gly Pro Ala Gly Ala 500 505 510 Ala Gly Arg Thr Arg Thr Ala Gly Tyr Leu Asp Ala Leu Leu Thr Val 515 520 525 Cys Leu Ala Arg Ala Gln His Gly Gln Ser Val 530 535 <210> SEQ ID NO 92 <211> LENGTH: 858 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 92 Met Gln His His His His His His Met Ser Asp Lys Ile Ile His Leu 5 10 15 Thr Asp Asp Ser Phe Asp Thr Asp Val Leu Lys Ala Asp Gly Ala Ile 20 25 30 Leu Val Asp Phe Trp Ala Glu Trp Cys Gly Pro Cys Lys Met Ile Ala 35 40 45 Pro Ile Leu Asp Glu Ile Ala Asp Glu Tyr Gln Gly Lys Leu Thr Val 50 55 60 Ala Lys Leu Asn Ile Asp Gln Asn Pro Gly Thr Ala Pro Lys Tyr Gly 65 70 75 80 Ile Arg Gly Ile Pro Thr Leu Leu Leu Phe Lys Asn Gly Glu Val Ala 85 90 95 Ala Thr Lys Val Gly Ala Leu Ser Lys Gly Gln Leu Lys Glu Phe Leu 100 105 110 Asp Ala Asn Leu Ala Gly Ser Gly Ser Gly His Met Gln His His His 115 120 125 His His His Val Ser Ile Glu Gly Arg Ala Ser Ser Gly Gly Ser Gly 130 135 140 Leu Val Pro Arg Gly Ser Ser Gly Ser Gly Asp Asp Asp Asp Lys Ser 145 150 155 160 Ser Arg His Ser Val Arg Gly His Ala Val Arg Arg Arg Arg Ala Ser 165 170 175 Thr Arg Ser His Ala Pro Ser Ala His Arg Ala Asp Ser Pro Val Glu 180 185 190 Asp Glu Pro Glu Gly Gly Gly Val Gly Leu Met Gly Tyr Leu Arg Ala 195 200 205 Val Phe Asn Val Asp Asp Asp Ser Glu Val Glu Ala Ala Gly Glu Met 210 215 220 Ala Ser Glu Glu Pro Pro Pro Arg Arg Arg Arg Glu Ala Arg Gly His 225 230 235 240 Pro Gly Ser Arg Arg Ala Ser Glu Ala Arg Ala Ala Ala Pro Pro Arg 245 250 255 Arg Ala Ser Phe Pro Arg Pro Arg Ser Val Thr Ala Arg Ser Gln Ser 260 265 270 Val Arg Gly Arg Arg Asp Ser Ala Ile Thr Arg Ala Pro Arg Gly Gly 275 280 285 Tyr Leu Gly Pro Met Asp Pro Arg Asp Val Leu Gly Arg Val Gly Gly 290 295 300 Ser Arg Val Val Pro Ser Pro Leu Phe Leu Asp Glu Leu Ser Tyr Glu 305 310 315 320 Glu Asp Asp Tyr Pro Ala Ala Val Ala His Asp Asp Gly Ala Gly Ala 325 330 335 Arg Pro Pro Ala Thr Val Glu Ile Leu Ala Gly Arg Val Ser Gly Pro 340 345 350 Glu Leu Gln Ala Ala Phe Pro Leu Asp Arg Leu Thr Pro Arg Val Ala 355 360 365 Ala Trp Asp Glu Ser Val Arg Ser Ala Leu Ala Leu Gly His Pro Ala 370 375 380 Gly Phe Tyr Pro Cys Pro Asp Ser Ala Phe Gly Leu Ser Arg Val Gly 385 390 395 400 Val Met His Phe Ala Ser Pro Ala Asp Pro Lys Val Phe Phe Arg Gln 405 410 415 Thr Leu Gln Gln Gly Glu Ala Leu Ala Trp Tyr Val Thr Gly Asp Ala 420 425 430 Ile Leu Asp Leu Thr Asp Arg Arg Ala Lys Thr Ser Pro Ser Arg Ala 435 440 445 Met Gly Phe Leu Val Asp Ala Ile Val Arg Val Ala Ile Asn Gly Trp 450 455 460 Val Cys Gly Thr Arg Leu His Thr Glu Gly Arg Gly Ser Glu Leu Asp 465 470 475 480 Asp Arg Ala Ala Glu Leu Arg Arg Gln Phe Ala Ser Leu Thr Ala Leu 485 490 495 Arg Pro Val Gly Ala Ala Ala Val Pro Leu Leu Ser Ala Gly Gly Ala 500 505 510 Ala Pro Pro His Pro Gly Pro Asp Ala Ala Val Phe Arg Ser Ser Leu 515 520 525 Gly Ser Leu Leu Tyr Trp Pro Gly Val Arg Ala Leu Leu Gly Arg Asp 530 535 540 Cys Arg Val Ala Ala Arg Tyr Ala Gly Arg Met Thr Tyr Ile Ala Thr 545 550 555 560 Gly Ala Leu Leu Ala Arg Phe Asn Pro Gly Ala Val Lys Cys Val Leu 565 570 575 Pro Arg Glu Ala Ala Phe Ala Gly Arg Val Leu Asp Val Leu Ala Val 580 585 590 Leu Ala Glu Gln Thr Val Gln Trp Leu Ser Val Val Val Gly Ala Arg 595 600 605 Leu His Pro His Ser Ala His Pro Ala Phe Ala Asp Val Glu Gln Glu 610 615 620 Ala Leu Phe Arg Ala Leu Pro Leu Gly Ser Pro Gly Val Val Ala Ala 625 630 635 640 Glu His Glu Ala Leu Gly Asp Thr Ala Ala Arg Arg Leu Leu Ala Thr 645 650 655 Ser Gly Leu Asn Ala Val Leu Gly Ala Ala Val Tyr Ala Leu His Thr 660 665 670 Ala Leu Ala Thr Val Thr Leu Lys Tyr Ala Leu Ala Cys Gly Asp Ala 675 680 685 Arg Arg Arg Arg Asp Asp Ala Ala Ala Ala Arg Ala Val Leu Ala Thr 690 695 700 Gly Leu Ile Leu Gln Arg Leu Leu Gly Leu Ala Asp Thr Val Val Ala 705 710 715 720 Cys Val Ala Leu Ala Ala Phe Asp Gly Gly Ser Thr Ala Pro Glu Val 725 730 735 Gly Thr Tyr Thr Pro Leu Arg Tyr Ala Cys Val Leu Arg Ala Thr Gln 740 745 750 Pro Leu Tyr Ala Arg Thr Thr Pro Ala Lys Phe Trp Ala Asp Val Arg 755 760 765 Ala Ala Ala Glu His Val Asp Leu Arg Pro Ala Ser Ser Ala Pro Arg 770 775 780 Ala Pro Val Ser Gly Thr Ala Asp Pro Ala Phe Leu Leu Glu Asp Leu 785 790 795 800 Ala Ala Phe Pro Pro Ala Pro Leu Asn Ser Glu Ser Val Leu Gly Pro 805 810 815 Arg Val Arg Val Val Asp Ile Met Ala Gln Phe Arg Lys Leu Leu Met 820 825 830 Gly Asp Glu Glu Thr Ala Ala Leu Arg Ala His Val Ser Gly Arg Arg 835 840 845 Ala Thr Gly Leu Gly Gly Pro Pro Arg Pro 850 855 <210> SEQ ID NO 93 <211> LENGTH: 311 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 93 Met Gln His His His His His His His Ser Val Arg Gly His Ala Val 5 10 15 Arg Arg Arg Arg Ala Ser Thr Arg Ser His Ala Pro Ser Ala His Arg 20 25 30 Ala Asp Ser Pro Val Glu Asp Glu Pro Glu Gly Gly Gly Val Gly Leu 35 40 45 Met Gly Tyr Leu Arg Ala Val Phe Asn Val Asp Asp Asp Ser Glu Val 50 55 60 Glu Ala Ala Gly Glu Met Ala Ser Glu Glu Pro Pro Pro Arg Arg Arg 65 70 75 80 Arg Glu Ala Arg Gly His Pro Gly Ser Arg Arg Ala Ser Glu Ala Arg 85 90 95 Ala Ala Ala Pro Pro Arg Arg Ala Ser Phe Pro Arg Pro Arg Ser Val 100 105 110 Thr Ala Arg Ser Gln Ser Val Arg Gly Arg Arg Asp Ser Ala Ile Thr 115 120 125 Arg Ala Pro Arg Gly Gly Tyr Leu Gly Pro Met Asp Pro Arg Asp Val 130 135 140 Leu Gly Arg Val Gly Gly Ser Arg Val Val Pro Ser Pro Leu Phe Leu 145 150 155 160 Asp Glu Leu Ser Tyr Glu Glu Asp Asp Tyr Pro Ala Ala Val Ala His 165 170 175 Asp Asp Gly Ala Gly Ala Arg Pro Pro Ala Thr Val Glu Ile Leu Ala 180 185 190 Gly Arg Val Ser Gly Pro Glu Leu Gln Ala Ala Phe Pro Leu Asp Arg 195 200 205 Leu Thr Pro Arg Val Ala Ala Trp Asp Glu Ser Val Arg Ser Ala Leu 210 215 220 Ala Leu Gly His Pro Ala Gly Phe Tyr Pro Cys Pro Asp Ser Ala Phe 225 230 235 240 Gly Leu Ser Arg Val Gly Val Met His Phe Ala Ser Pro Ala Asp Pro 245 250 255 Lys Val Phe Phe Arg Gln Thr Leu Gln Gln Gly Glu Ala Leu Ala Trp 260 265 270 Tyr Val Thr Gly Asp Ala Ile Leu Asp Leu Thr Asp Arg Arg Ala Lys 275 280 285 Thr Ser Pro Ser Arg Ala Met Gly Phe Leu Val Asp Ala Ile Val Arg 290 295 300 Val Ala Ile Asn Gly Trp Val 305 310 <210> SEQ ID NO 94 <211> LENGTH: 704 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 94 Met Gln His His His His His His His Ser Val Arg Gly His Ala Val 5 10 15 Arg Arg Arg Arg Ala Ser Thr Arg Ser His Ala Pro Ser Ala His Arg 20 25 30 Ala Asp Ser Pro Val Glu Asp Glu Pro Glu Gly Gly Gly Val Gly Leu 35 40 45 Met Gly Tyr Leu Arg Ala Val Phe Asn Val Asp Asp Asp Ser Glu Val 50 55 60 Glu Ala Ala Gly Glu Met Ala Ser Glu Glu Pro Pro Pro Arg Arg Arg 65 70 75 80 Arg Glu Ala Arg Gly His Pro Gly Ser Arg Arg Ala Ser Glu Ala Arg 85 90 95 Ala Ala Ala Pro Pro Arg Arg Ala Ser Phe Pro Arg Pro Arg Ser Val 100 105 110 Thr Ala Arg Ser Gln Ser Val Arg Gly Arg Arg Asp Ser Ala Ile Thr 115 120 125 Arg Ala Pro Arg Gly Gly Tyr Leu Gly Pro Met Asp Pro Arg Asp Val 130 135 140 Leu Gly Arg Val Gly Gly Ser Arg Val Val Pro Ser Pro Leu Phe Leu 145 150 155 160 Asp Glu Leu Ser Tyr Glu Glu Asp Asp Tyr Pro Ala Ala Val Ala His 165 170 175 Asp Asp Gly Ala Gly Ala Arg Pro Pro Ala Thr Val Glu Ile Leu Ala 180 185 190 Gly Arg Val Ser Gly Pro Glu Leu Gln Ala Ala Phe Pro Leu Asp Arg 195 200 205 Leu Thr Pro Arg Val Ala Ala Trp Asp Glu Ser Val Arg Ser Ala Leu 210 215 220 Ala Leu Gly His Pro Ala Gly Phe Tyr Pro Cys Pro Asp Ser Ala Phe 225 230 235 240 Gly Leu Ser Arg Val Gly Val Met His Phe Ala Ser Pro Ala Asp Pro 245 250 255 Lys Val Phe Phe Arg Gln Thr Leu Gln Gln Gly Glu Ala Leu Ala Trp 260 265 270 Tyr Val Thr Gly Asp Ala Ile Leu Asp Leu Thr Asp Arg Arg Ala Lys 275 280 285 Thr Ser Pro Ser Arg Ala Met Gly Phe Leu Val Asp Ala Ile Val Arg 290 295 300 Val Ala Ile Asn Gly Trp Val Cys Gly Thr Arg Leu His Thr Glu Gly 305 310 315 320 Arg Gly Ser Glu Leu Asp Asp Arg Ala Ala Glu Leu Arg Arg Gln Phe 325 330 335 Ala Ser Leu Thr Ala Leu Arg Pro Val Gly Ala Ala Ala Val Pro Leu 340 345 350 Leu Ser Ala Gly Gly Ala Ala Pro Pro His Pro Gly Pro Asp Ala Ala 355 360 365 Val Phe Arg Ser Ser Leu Gly Ser Leu Leu Tyr Trp Pro Gly Val Arg 370 375 380 Ala Leu Leu Gly Arg Asp Cys Arg Val Ala Ala Arg Tyr Ala Gly Arg 385 390 395 400 Met Thr Tyr Ile Ala Thr Gly Ala Leu Leu Ala Arg Phe Asn Pro Gly 405 410 415 Ala Val Lys Cys Val Leu Pro Arg Glu Ala Ala Phe Ala Gly Arg Val 420 425 430 Leu Asp Val Leu Ala Val Leu Ala Glu Gln Thr Val Gln Trp Leu Ser 435 440 445 Val Val Val Gly Ala Arg Leu His Pro His Ser Ala His Pro Ala Phe 450 455 460 Ala Asp Val Glu Gln Glu Ala Leu Phe Arg Ala Leu Pro Leu Gly Ser 465 470 475 480 Pro Gly Val Val Ala Ala Glu His Glu Ala Leu Gly Asp Thr Ala Ala 485 490 495 Arg Arg Leu Leu Ala Thr Ser Gly Leu Asn Ala Val Leu Gly Ala Ala 500 505 510 Val Tyr Ala Leu His Thr Ala Leu Ala Thr Val Thr Leu Lys Tyr Ala 515 520 525 Leu Ala Cys Gly Asp Ala Arg Arg Arg Arg Asp Asp Ala Ala Ala Ala 530 535 540 Arg Ala Val Leu Ala Thr Gly Leu Ile Leu Gln Arg Leu Leu Gly Leu 545 550 555 560 Ala Asp Thr Val Val Ala Cys Val Ala Leu Ala Ala Phe Asp Gly Gly 565 570 575 Ser Thr Ala Pro Glu Val Gly Thr Tyr Thr Pro Leu Arg Tyr Ala Cys 580 585 590 Val Leu Arg Ala Thr Gln Pro Leu Tyr Ala Arg Thr Thr Pro Ala Lys 595 600 605 Phe Trp Ala Asp Val Arg Ala Ala Ala Glu His Val Asp Leu Arg Pro 610 615 620 Ala Ser Ser Ala Pro Arg Ala Pro Val Ser Gly Thr Ala Asp Pro Ala 625 630 635 640 Phe Leu Leu Glu Asp Leu Ala Ala Phe Pro Pro Ala Pro Leu Asn Ser 645 650 655 Glu Ser Val Leu Gly Pro Arg Val Arg Val Val Asp Ile Met Ala Gln 660 665 670 Phe Arg Lys Leu Leu Met Gly Asp Glu Glu Thr Ala Ala Leu Arg Ala 675 680 685 His Val Ser Gly Arg Arg Ala Thr Gly Leu Gly Gly Pro Pro Arg Pro 690 695 700 <210> SEQ ID NO 95 <211> LENGTH: 1381 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 95 Met Gln His His His His His His Ala Ala Pro Ala Arg Asp Pro Pro 5 10 15 Gly Tyr Arg Tyr Ala Ala Ala Met Val Pro Thr Gly Ser Ile Leu Ser 20 25 30 Thr Ile Glu Val Ala Ser His Arg Arg Leu Phe Asp Phe Phe Ala Arg 35 40 45 Val Arg Ser Asp Glu Asn Ser Leu Tyr Asp Val Glu Phe Asp Ala Leu 50 55 60 Leu Gly Ser Tyr Cys Asn Thr Leu Ser Leu Val Arg Phe Leu Glu Leu 65 70 75 80 Gly Leu Ser Val Ala Cys Val Cys Thr Lys Phe Pro Glu Leu Ala Tyr 85 90 95 Met Asn Glu Gly Arg Val Gln Phe Glu Val His Gln Pro Leu Ile Ala 100 105 110 Arg Asp Gly Pro His Pro Val Glu Gln Pro Val His Asn Tyr Met Thr 115 120 125 Lys Val Ile Asp Arg Arg Ala Leu Asn Ala Ala Phe Ser Leu Ala Thr 130 135 140 Glu Ala Ile Ala Leu Leu Thr Gly Glu Ala Leu Asp Gly Thr Gly Ile 145 150 155 160 Ser Leu His Arg Gln Leu Arg Ala Ile Gln Gln Leu Ala Arg Asn Val 165 170 175 Gln Ala Val Leu Gly Ala Phe Glu Arg Gly Thr Ala Asp Gln Met Leu 180 185 190 His Val Leu Leu Glu Lys Ala Pro Pro Leu Ala Leu Leu Leu Pro Met 195 200 205 Gln Arg Tyr Leu Asp Asn Gly Arg Leu Ala Thr Arg Val Ala Arg Ala 210 215 220 Thr Leu Val Ala Glu Leu Lys Arg Ser Phe Cys Asp Thr Ser Phe Phe 225 230 235 240 Leu Gly Lys Ala Gly His Arg Arg Glu Ala Ile Glu Ala Trp Leu Val 245 250 255 Asp Leu Thr Thr Ala Thr Gln Pro Ser Val Ala Val Pro Arg Leu Thr 260 265 270 His Ala Asp Thr Arg Gly Arg Pro Val Asp Gly Val Leu Val Thr Thr 275 280 285 Ala Ala Ile Lys Gln Arg Leu Leu Gln Ser Phe Leu Lys Val Glu Asp 290 295 300 Thr Glu Ala Asp Val Pro Val Thr Tyr Gly Glu Met Val Leu Asn Gly 305 310 315 320 Ala Asn Leu Val Thr Ala Leu Val Met Gly Lys Ala Val Arg Ser Leu 325 330 335 Asp Asp Val Gly Arg His Leu Leu Glu Met Gln Glu Glu Gln Leu Glu 340 345 350 Ala Asn Arg Glu Thr Leu Asp Glu Leu Glu Ser Ala Pro Gln Thr Thr 355 360 365 Arg Val Arg Ala Asp Leu Val Ala Ile Gly Asp Arg Leu Val Phe Leu 370 375 380 Glu Ala Leu Glu Lys Arg Ile Tyr Ala Ala Thr Asn Val Pro Tyr Pro 385 390 395 400 Leu Val Gly Ala Met Asp Leu Thr Phe Val Leu Pro Leu Gly Leu Phe 405 410 415 Asn Pro Ala Met Glu Arg Phe Ala Ala His Ala Gly Asp Leu Val Pro 420 425 430 Ala Pro Gly His Pro Glu Pro Arg Ala Phe Pro Pro Arg Gln Leu Phe 435 440 445 Phe Trp Gly Lys Asp His Gln Val Leu Arg Leu Ser Met Glu Asn Ala 450 455 460 Val Gly Thr Val Cys His Pro Ser Leu Met Asn Ile Asp Ala Ala Val 465 470 475 480 Gly Gly Val Asn His Ala Pro Val Glu Ala Ala Asn Pro Tyr Gly Ala 485 490 495 Tyr Val Ala Ala Pro Ala Gly Pro Gly Ala Asp Met Gln Gln Arg Phe 500 505 510 Leu Asn Ala Trp Arg Gln Arg Leu Ala His Gly Arg Val Arg Trp Val 515 520 525 Ala Glu Cys Gln Met Thr Ala Glu Gln Phe Met Gln Pro Asp Asn Ala 530 535 540 Asn Leu Ala Leu Glu Leu His Pro Ala Phe Asp Phe Phe Ala Gly Val 545 550 555 560 Ala Asp Val Glu Leu Pro Gly Gly Glu Val Pro Pro Ala Gly Pro Gly 565 570 575 Ala Ile Gln Ala Thr Trp Arg Val Val Asn Gly Asn Leu Pro Leu Ala 580 585 590 Leu Cys Pro Val Ala Phe Arg Asp Ala Arg Gly Leu Glu Leu Gly Val 595 600 605 Gly Arg His Ala Met Ala Pro Ala Thr Ile Ala Ala Val Arg Gly Ala 610 615 620 Phe Glu Asp Arg Ser Tyr Pro Ala Val Phe Tyr Leu Leu Gln Ala Ala 625 630 635 640 Ile His Gly Ser Glu His Val Phe Cys Ala Leu Ala Arg Leu Val Thr 645 650 655 Gln Cys Ile Thr Ser Tyr Trp Asn Asn Thr Arg Cys Ala Ala Phe Val 660 665 670 Asn Asp Tyr Ser Leu Val Ser Tyr Ile Val Thr Tyr Leu Gly Gly Asp 675 680 685 Leu Pro Glu Glu Cys Met Ala Val Tyr Arg Asp Leu Val Ala His Val 690 695 700 Glu Ala Leu Ala Gln Leu Val Asp Asp Phe Thr Leu Pro Gly Pro Glu 705 710 715 720 Leu Gly Gly Gln Ala Gln Ala Glu Leu Asn His Leu Met Arg Asp Pro 725 730 735 Ala Leu Leu Pro Pro Leu Val Trp Asp Cys Asp Gly Leu Met Arg His 740 745 750 Ala Ala Leu Asp Arg His Arg Asp Cys Arg Ile Asp Ala Gly Gly His 755 760 765 Glu Pro Val Tyr Ala Ala Ala Cys Asn Val Ala Thr Ala Asp Phe Asn 770 775 780 Arg Asn Asp Gly Arg Leu Leu His Asn Thr Gln Ala Arg Ala Val Asp 785 790 795 800 Ala Ala Asp Asp Arg Pro His Arg Pro Ala Asp Trp Thr Val His His 805 810 815 Lys Ile Tyr Tyr Tyr Val Leu Val Pro Ala Phe Ser Arg Gly Arg Cys 820 825 830 Cys Thr Ala Gly Val Arg Phe Asp Arg Val Tyr Ala Thr Leu Gln Asn 835 840 845 Met Val Val Pro Glu Ile Ala Pro Gly Glu Glu Cys Pro Ser Asp Pro 850 855 860 Val Thr Asp Pro Ala His Pro Leu His Pro Ala Asn Leu Val Ala Asn 865 870 875 880 Thr Val Asn Ala Met Phe His Asn Gly Arg Val Val Val Asp Gly Pro 885 890 895 Ala Met Leu Thr Leu Gln Val Leu Ala His Asn Met Ala Glu Arg Thr 900 905 910 Thr Ala Leu Leu Cys Ser Ala Ala Pro Asp Ala Gly Ala Asn Thr Ala 915 920 925 Ser Thr Ala Asn Met Arg Ile Phe Asp Gly Ala Leu His Ala Gly Val 930 935 940 Leu Leu Met Ala Pro Gln His Leu Asp His Thr Ile Gln Asn Gly Glu 945 950 955 960 Tyr Phe Tyr Val Leu Pro Val His Ala Leu Phe Ala Gly Ala Asp His 965 970 975 Val Ala Asn Ala Pro Asn Phe Pro Pro Ala Leu Arg Asp Leu Ala Arg 980 985 990 His Val Pro Leu Val Pro Pro Ala Leu Gly Ala Asn Tyr Phe Ser Ser 995 1000 1005 Ile Arg Gln Pro Val Val Gln His Ala Arg Glu Ser Ala Ala Gly Glu 1010 1015 1020 Asn Ala Leu Thr Tyr Ala Leu Met Ala Gly Tyr Phe Lys Met Ser Pro 1025 1030 1035 1040 Val Ala Leu Tyr His Gln Leu Lys Thr Gly Leu His Pro Gly Phe Gly 1045 1050 1055 Phe Thr Val Val Arg Gln Asp Arg Phe Val Thr Glu Asn Val Leu Phe 1060 1065 1070 Ser Glu Arg Ala Ser Glu Ala Tyr Phe Leu Gly Gln Leu Gln Val Ala 1075 1080 1085 Arg His Glu Thr Gly Gly Gly Val Ser Phe Thr Leu Thr Gln Pro Arg 1090 1095 1100 Gly Asn Val Asp Leu Gly Val Gly Tyr Thr Ala Val Ala Ala Thr Ala 1105 1110 1115 1120 Thr Val Arg Asn Pro Val Thr Asp Met Gly Asn Leu Pro Gln Asn Phe 1125 1130 1135 Tyr Leu Gly Arg Gly Ala Pro Pro Leu Leu Asp Asn Ala Ala Ala Val 1140 1145 1150 Tyr Leu Arg Asn Ala Val Val Ala Gly Asn Arg Leu Gly Pro Ala Gln 1155 1160 1165 Pro Leu Pro Val Phe Gly Cys Ala Gln Val Pro Arg Arg Ala Gly Met 1170 1175 1180 Asp His Gly Gln Asp Ala Val Cys Glu Phe Ile Ala Thr Pro Val Ala 1185 1190 1195 1200 Thr Asp Ile Asn Tyr Phe Arg Arg Pro Cys Asn Pro Arg Gly Arg Ala 1205 1210 1215 Ala Gly Gly Val Tyr Ala Gly Asp Lys Glu Gly Asp Val Ile Ala Leu 1220 1225 1230 Met Tyr Asp His Gly Gln Ser Asp Pro Ala Arg Pro Phe Ala Ala Thr 1235 1240 1245 Ala Asn Pro Trp Ala Ser Gln Arg Phe Ser Tyr Gly Asp Leu Leu Tyr 1250 1255 1260 Asn Gly Ala Tyr His Leu Asn Gly Ala Ser Pro Val Leu Ser Pro Cys 1265 1270 1275 1280 Phe Lys Phe Phe Thr Ala Ala Asp Ile Thr Ala Lys His Arg Cys Leu 1285 1290 1295 Glu Arg Leu Ile Val Glu Thr Gly Ser Ala Val Ser Thr Ala Thr Ala 1300 1305 1310 Ala Ser Asp Val Gln Phe Lys Arg Pro Pro Gly Cys Arg Glu Leu Val 1315 1320 1325 Glu Asp Pro Cys Gly Leu Phe Gln Glu Ala Tyr Pro Ile Thr Cys Ala 1330 1335 1340 Ser Asp Pro Ala Leu Leu Arg Ser Ala Arg Asp Gly Glu Ala His Ala 1345 1350 1355 1360 Arg Glu Thr His Phe Thr Gln Tyr Leu Ile Tyr Asp Ala Ser Pro Leu 1365 1370 1375 Lys Gly Leu Ser Leu 1380 <210> SEQ ID NO 96 <211> LENGTH: 377 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 96 Met Gln His His His His His His His Ser Gln Trp Gly Pro Arg Ala 5 10 15 Ile Leu Val Gln Thr Asp Ser Thr Asn Arg Asn Ala Asp Gly Asp Trp 20 25 30 Gln Ala Ala Val Ala Ile Arg Gly Gly Gly Val Val Gln Leu Asn Met 35 40 45 Val Asn Lys Arg Ala Val Asp Phe Thr Pro Ala Glu Cys Gly Asp Ser 50 55 60 Glu Trp Ala Val Gly Arg Val Ser Leu Gly Leu Arg Met Ala Met Pro 65 70 75 80 Arg Asp Phe Cys Ala Ile Ile His Ala Pro Ala Val Ser Gly Pro Gly 85 90 95 Pro His Val Met Leu Gly Leu Val Asp Ser Gly Tyr Arg Gly Thr Val 100 105 110 Leu Ala Val Val Val Ala Pro Asn Gly Thr Arg Gly Phe Ala Pro Gly 115 120 125 Ala Leu Arg Val Asp Val Thr Phe Leu Asp Ile Arg Ala Thr Pro Pro 130 135 140 Thr Leu Thr Glu Pro Ser Ser Leu His Arg Phe Pro Gln Leu Ala Pro 145 150 155 160 Ser Pro Leu Ala Gly Leu Arg Glu Asp Pro Trp Leu Asp Gly Ala Leu 165 170 175 Ala Thr Ala Gly Gly Ala Val Ala Leu Pro Ala Arg Arg Arg Gly Gly 180 185 190 Ser Leu Val Tyr Ala Gly Glu Leu Thr Gln Val Thr Thr Glu His Gly 195 200 205 Asp Cys Val His Glu Ala Pro Ala Phe Leu Pro Lys Arg Glu Glu Asp 210 215 220 Ala Gly Phe Asp Ile Leu Ile His Arg Ala Val Thr Val Pro Ala Asn 225 230 235 240 Gly Ala Thr Val Ile Gln Pro Ser Leu Arg Val Leu Arg Ala Ala Asp 245 250 255 Gly Pro Glu Ala Cys Tyr Val Leu Gly Arg Ser Ser Leu Asn Ala Arg 260 265 270 Gly Leu Leu Val Met Pro Thr Arg Trp Pro Ser Gly His Ala Cys Ala 275 280 285 Phe Val Val Cys Asn Leu Thr Gly Val Pro Val Thr Leu Gln Ala Gly 290 295 300 Ser Lys Val Ala Gln Leu Leu Val Ala Gly Thr His Ala Leu Pro Trp 305 310 315 320 Ile Pro Pro Asp Asn Ile His Glu Asp Gly Ala Phe Arg Ala Tyr Pro 325 330 335 Arg Gly Val Pro Asp Ala Thr Ala Thr Pro Arg Asp Pro Pro Ile Leu 340 345 350 Val Phe Thr Asn Glu Phe Asp Ala Asp Ala Pro Pro Ser Lys Arg Gly 355 360 365 Ala Gly Gly Phe Gly Ser Thr Gly Ile 370 375 <210> SEQ ID NO 97 <211> LENGTH: 308 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 97 Met Gln His His His His His His His Thr Ser Arg Arg Ser Val Lys 5 10 15 Ser Cys Pro Arg Glu Ala Pro Arg Gly Thr His Glu Glu Leu Tyr Tyr 20 25 30 Gly Pro Val Ser Pro Ala Asp Pro Glu Ser Pro Arg Asp Asp Phe Arg 35 40 45 Arg Gly Ala Gly Pro Met Arg Ala Arg Pro Arg Gly Glu Val Arg Phe 50 55 60 Leu His Tyr Asp Glu Ala Gly Tyr Ala Leu Tyr Arg Asp Ser Ser Ser 65 70 75 80 Asp Asp Asp Glu Ser Arg Asp Thr Ala Arg Pro Arg Arg Ser Ala Ser 85 90 95 Val Ala Gly Ser His Gly Pro Gly Pro Ala Arg Ala Pro Pro Pro Pro 100 105 110 Gly Gly Pro Val Gly Ala Gly Gly Arg Ser His Ala Pro Pro Ala Arg 115 120 125 Thr Pro Lys Met Thr Arg Gly Ala Pro Lys Ala Pro Ala Thr Pro Ala 130 135 140 Thr Asp Pro Ala Arg Gly Arg Arg Pro Ala Gln Ala Asp Ser Ala Val 145 150 155 160 Leu Leu Asp Ala Pro Ala Pro Thr Ala Ser Gly Arg Thr Lys Thr Pro 165 170 175 Ala Gln Gly Leu Ala Lys Lys Leu His Phe Ser Thr Ala Pro Pro Ser 180 185 190 Pro Thr Ala Pro Trp Thr Pro Arg Val Ala Gly Phe Asn Lys Arg Val 195 200 205 Phe Cys Ala Ala Val Gly Arg Leu Ala Ala Thr His Ala Arg Leu Ala 210 215 220 Ala Val Gln Leu Trp Asp Met Ser Arg Pro His Thr Asp Glu Asp Leu 225 230 235 240 Asn Glu Leu Leu Asp Leu Thr Thr Ile Arg Val Thr Val Cys Glu Gly 245 250 255 Lys Asn Leu Leu Gln Arg Ala Asn Glu Leu Val Asn Pro Asp Ala Ala 260 265 270 Gln Asp Val Asp Ala Thr Ala Ala Ala Arg Gly Arg Pro Ala Gly Arg 275 280 285 Ala Ala Ala Thr Ala Arg Ala Pro Ala Arg Ser Ala Ser Arg Pro Arg 290 295 300 Arg Pro Leu Glu 305 <210> SEQ ID NO 98 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 98 gagctcagct atgccaccac c 21 <210> SEQ ID NO 99 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 99 cggcgaattc attagtagag gcggtggaaa aag 33 <210> SEQ ID NO 100 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 100 cacgccgccg caccccaggc ggac 24 <210> SEQ ID NO 101 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 101 cggcgaattc attagtagag gcggtggaaa aag 33 <210> SEQ ID NO 102 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 102 cacacctctc gccgctccgt caagtc 26 <210> SEQ ID NO 103 <211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 103 cataagaatt cactactcga gggggcggcg gggacg 36 <210> SEQ ID NO 104 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 104 cacagtcagt gggggcccag ggcgatcc 28 <210> SEQ ID NO 105 <211> LENGTH: 35 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 105 cctagaattc actagatgcc agtggagcca aaccc 35 <210> SEQ ID NO 106 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 106 gccgctcctg cccgcgaccc ccc 23 <210> SEQ ID NO 107 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 107 ccagaattca ttacagagac aggcccttta gc 32 <210> SEQ ID NO 108 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 108 cactccgtgg cgcgggcatg ccg 23 <210> SEQ ID NO 109 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 109 ccgttagaat tcactatggg cgtggcgggc c 31 <210> SEQ ID NO 110 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 110 cactccgtgc gcgggcatgc cg 22 <210> SEQ ID NO 111 <211> LENGTH: 38 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 111 catagaattc atcacgcgcg ggaggggctg gtttttgc 38 <210> SEQ ID NO 112 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 112 gacacggtgg tcgcgtgcgt ggc 23 <210> SEQ ID NO 113 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 113 ccgttagaat tcactatggg cgtggcgggc c 31 <210> SEQ ID NO 114 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 114 cactccgtgc gcgggcatgc cg 22 <210> SEQ ID NO 115 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 115 cgtatgaatt catcagaccc acccgttg 28 <210> SEQ ID NO 116 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 116 gtgctggcga cggggctcat cc 22 <210> SEQ ID NO 117 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 117 ccgttagaat tcactatggg cgtggcgggc c 31 <210> SEQ ID NO 118 <211> LENGTH: 783 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 118 atggctcgcg gggccgggtt ggtgtttttt gttggagttt gggtcgtatc gtgcctggcg 60 gcagcaccca gaacgtcctg gaaacgggta acctcgggcg aggacgtggt gttgcttccg 120 gcgcccgcgg aacgcacccg ggcccacaaa ctactatggg ccgcggaacc cctggatgcc 180 tgcggtcccc tgcgcccgtc gtgggtggcg ctgtggcccc cccgacgggt gctcgagacg 240 gtcgtggatg cggcgtgcat gcgcgccccg gaaccgctcg ccatagcata cagtcccccg 300 ttccccgcgg gcgacgaggg actgtattcg gagttggcgt ggcgcgatcg cgtagccgtg 360 gtcaacgaga gtctggtcat ctacggggcc ctggagacgg acagcggtct gtacaccctg 420 tccgtggtcg gcctaagcga cgaggcgcgc caagtggcgt cggtggttct ggtcgtggag 480 cccgcccctg tgccgacccc gacccccgac gactacgacg aagaagacga cgcgggcgtg 540 agcgaacgca cgccggtcag cgttcccccc ccaacccccc ccccgtcgtc cccccgtcgc 600 ccccccgacg caccctcgtg ttatccccga ggtgtcccac gtgcgcgggg taacggtcca 660 tatggagacc ccggaggcca ttctgtttgc ccccggggag acgtttggga cgaacgtctc 720 catccacgcc attgcccacg acgacggtcc gtacgccatg gacgtcgtct ggatgcggtt 780 tga 783 <210> SEQ ID NO 119 <211> LENGTH: 1638 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 119 atggctcgcg gggccgggtt ggtgtttttt gttggagttt gggtcgtatc gtgcctggcg 60 gcagcaccca gaacgtcctg gaaacgggta acctcgggcg aggacgtggt gttgcttccg 120 gcgcccgcgg aacgcacccg ggcccacaaa ctactgtggg ccgcggaacc cctggatgcc 180 tgcggtcccc tgcgcccgtc gtgggtggcg ctgtggcccc cccgacgggt gctcgagacg 240 gtcgtggatg cggcgtgcat gcgcgccccg gaaccgctcg ccatagcata cagtcccccg 300 ttccccgcgg gcgacgaggg actgtattcg gagttggcgt ggcgcgatcg cgtagccgtg 360 gtcaacgaga gtctggtcat ctacggggcc ctggagacgg acagcggtct gtacaccctg 420 tccgtggtcg gcctaagcga cgaggcgcgc caagtggcgt cggtggttct ggtcgtggag 480 cccgcccctg tgccgacccc gacccccgac gactacgacg aagaagacga cgcgggcgtg 540 acgaacgcac gccggtcagc gttccccccc caaccccccc cccgtcgtcc ccccgtcgcc 600 cccccgacgc accctcgtgt tatccccgag gtgtcccacg tgcgcggggt aacggtccat 660 atggagaccc tggaggccat tctgtttgcc cccggggaga cgtttgggac gaacgtctcc 720 atccacgcca ttgcccacga cgacggtccg tacgccatgg acgtcgtctg gatgcggttt 780 gacgtgccgt cctcgtgcgc cgatatgcgg atctacgaag cttgtctgta tcacccgcag 840 cttccagagt gtctatctcc ggccgacgcg ccgtgcgccg taagttcctg ggcgtaccgc 900 ctggcggtcc gcagctacgc cggctgttcc aggactacgc ccccgccgcg atgttttgcc 960 gaggctcgca tggaaccggt cccggggttg gcgtggctgg cctccaccgt caatctggaa 1020 ttccagcacg cctcccccca gcacgccggc ctctacctgt gcgtggtgta cgtggacgat 1080 catatccacg cctggggcca catgaccatc agcaccgcgg cgcagtaccg gaacgcggtg 1140 gtggaacagc acctccccca gcgccagccc gagcccgtcg agcccacccg cccgcacgtg 1200 agagcccccc atcccgcgcc ctccgcgcgc ggcccgctgc gcctcggggc ggtgctgggg 1260 gcggccctgt tgctggccgc cctcgggctg tccgcgtggg cgtgcatgac ctgctggcgc 1320 aggcgctcct ggcgggcggt taaaagccgg gcctcggcga cgggccccac ttacattcgc 1380 gtggcggaca gcgagctgta cgcggactgg agttcggaca gcgaggggga gcgcgacggg 1440 tccctgtggc aggaccctcc ggagagaccc gactctccct ccacaaatgg atccggcttt 1500 gagatcttat caccaacggc tccgtctgta tacccccata gcgaggggcg taaatctcgc 1560 cgcccgctca ccacctttgg ttcgggaagc ccgggccgtc gtcactccca ggcctcctat 1620 ccgtccgtcc tctggtaa 1638 <210> SEQ ID NO 120 <211> LENGTH: 260 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 120 Met Ala Arg Gly Ala Gly Leu Val Phe Phe Val Gly Val Trp Val Val 5 10 15 Ser Cys Leu Ala Ala Ala Pro Arg Thr Ser Trp Lys Arg Val Thr Ser 20 25 30 Gly Glu Asp Val Val Leu Leu Pro Ala Pro Ala Glu Arg Thr Arg Ala 35 40 45 His Lys Leu Leu Trp Ala Ala Glu Pro Leu Asp Ala Cys Gly Pro Leu 50 55 60 Arg Pro Ser Trp Val Ala Leu Trp Pro Pro Arg Arg Val Leu Glu Thr 65 70 75 80 Val Val Asp Ala Ala Cys Met Arg Ala Pro Glu Pro Leu Ala Ile Ala 85 90 95 Tyr Ser Pro Pro Phe Pro Ala Gly Asp Glu Gly Leu Tyr Ser Glu Leu 100 105 110 Ala Trp Arg Asp Arg Val Ala Val Val Asn Glu Ser Leu Val Ile Tyr 115 120 125 Gly Ala Leu Glu Thr Asp Ser Gly Leu Tyr Thr Leu Ser Val Val Gly 130 135 140 Leu Ser Asp Glu Ala Arg Gln Val Ala Ser Val Val Leu Val Val Glu 145 150 155 160 Pro Ala Pro Val Pro Thr Pro Thr Pro Asp Asp Tyr Asp Glu Glu Asp 165 170 175 Asp Ala Gly Val Ser Glu Arg Thr Pro Val Ser Val Pro Pro Pro Thr 180 185 190 Pro Pro Pro Ser Ser Pro Arg Arg Pro Pro Asp Ala Pro Ser Cys Tyr 195 200 205 Pro Arg Gly Val Pro Arg Ala Arg Gly Asn Gly Pro Tyr Gly Asp Pro 210 215 220 Gly Gly His Ser Val Cys Pro Arg Gly Asp Val Trp Asp Glu Arg Leu 225 230 235 240 His Pro Arg His Cys Pro Arg Arg Arg Ser Val Arg His Gly Arg Arg 245 250 255 Leu Asp Ala Val 260 <210> SEQ ID NO 121 <211> LENGTH: 545 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 121 Met Ala Arg Gly Ala Gly Leu Val Phe Phe Val Gly Val Trp Val Val 5 10 15 Ser Cys Leu Ala Ala Ala Pro Arg Thr Ser Trp Lys Arg Val Thr Ser 20 25 30 Gly Glu Asp Val Val Leu Leu Pro Ala Pro Ala Glu Arg Thr Arg Ala 35 40 45 His Lys Leu Leu Trp Ala Ala Glu Pro Leu Asp Ala Cys Gly Pro Leu 50 55 60 Arg Pro Ser Trp Val Ala Leu Trp Pro Pro Arg Arg Val Leu Glu Thr 65 70 75 80 Val Val Asp Ala Ala Cys Met Arg Ala Pro Glu Pro Leu Ala Ile Ala 85 90 95 Tyr Ser Pro Pro Phe Pro Ala Gly Asp Glu Gly Leu Tyr Ser Glu Leu 100 105 110 Ala Trp Arg Asp Arg Val Ala Val Val Asn Glu Ser Leu Val Ile Tyr 115 120 125 Gly Ala Leu Glu Thr Asp Ser Gly Leu Tyr Thr Leu Ser Val Val Gly 130 135 140 Leu Ser Asp Glu Ala Arg Gln Val Ala Ser Val Val Leu Val Val Glu 145 150 155 160 Pro Ala Pro Val Pro Thr Pro Thr Pro Asp Asp Tyr Asp Glu Glu Asp 165 170 175 Asp Ala Gly Val Thr Asn Ala Arg Arg Ser Ala Phe Pro Pro Gln Pro 180 185 190 Pro Pro Arg Arg Pro Pro Val Ala Pro Pro Thr His Pro Arg Val Ile 195 200 205 Pro Glu Val Ser His Val Arg Gly Val Thr Val His Met Glu Thr Leu 210 215 220 Glu Ala Ile Leu Phe Ala Pro Gly Glu Thr Phe Gly Thr Asn Val Ser 225 230 235 240 Ile His Ala Ile Ala His Asp Asp Gly Pro Tyr Ala Met Asp Val Val 245 250 255 Trp Met Arg Phe Asp Val Pro Ser Ser Cys Ala Asp Met Arg Ile Tyr 260 265 270 Glu Ala Cys Leu Tyr His Pro Gln Leu Pro Glu Cys Leu Ser Pro Ala 275 280 285 Asp Ala Pro Cys Ala Val Ser Ser Trp Ala Tyr Arg Leu Ala Val Arg 290 295 300 Ser Tyr Ala Gly Cys Ser Arg Thr Thr Pro Pro Pro Arg Cys Phe Ala 305 310 315 320 Glu Ala Arg Met Glu Pro Val Pro Gly Leu Ala Trp Leu Ala Ser Thr 325 330 335 Val Asn Leu Glu Phe Gln His Ala Ser Pro Gln His Ala Gly Leu Tyr 340 345 350 Leu Cys Val Val Tyr Val Asp Asp His Ile His Ala Trp Gly His Met 355 360 365 Thr Ile Ser Thr Ala Ala Gln Tyr Arg Asn Ala Val Val Glu Gln His 370 375 380 Leu Pro Gln Arg Gln Pro Glu Pro Val Glu Pro Thr Arg Pro His Val 385 390 395 400 Arg Ala Pro His Pro Ala Pro Ser Ala Arg Gly Pro Leu Arg Leu Gly 405 410 415 Ala Val Leu Gly Ala Ala Leu Leu Leu Ala Ala Leu Gly Leu Ser Ala 420 425 430 Trp Ala Cys Met Thr Cys Trp Arg Arg Arg Ser Trp Arg Ala Val Lys 435 440 445 Ser Arg Ala Ser Ala Thr Gly Pro Thr Tyr Ile Arg Val Ala Asp Ser 450 455 460 Glu Leu Tyr Ala Asp Trp Ser Ser Asp Ser Glu Gly Glu Arg Asp Gly 465 470 475 480 Ser Leu Trp Gln Asp Pro Pro Glu Arg Pro Asp Ser Pro Ser Thr Asn 485 490 495 Gly Ser Gly Phe Glu Ile Leu Ser Pro Thr Ala Pro Ser Val Tyr Pro 500 505 510 His Ser Glu Gly Arg Lys Ser Arg Arg Pro Leu Thr Thr Phe Gly Ser 515 520 525 Gly Ser Pro Gly Arg Arg His Ser Gln Ala Ser Tyr Pro Ser Val Leu 530 535 540 Trp 545 <210> SEQ ID NO 122 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 122 Val Gly Ala Ala Ala Val Pro Leu Leu Ser Ala Gly Gly Ala Ala 1 5 10 15 <210> SEQ ID NO 123 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 123 Pro His Pro Gly Pro Asp Ala Ala Val Phe Arg Ser Ser Leu Gly 1 5 10 15 <210> SEQ ID NO 124 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 124 Met Thr Tyr Ile Ala Thr Gly Ala Leu Leu Ala 1 5 10 <210> SEQ ID NO 125 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 125 Glu Ala Ala Phe Ala Gly Arg Val Leu Asp Val Leu Ala Val Leu 1 5 10 15 <210> SEQ ID NO 126 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 126 Ala Arg Leu His Pro His Ser Ala His Pro Ala Phe Ala Asp Val 1 5 10 15 <210> SEQ ID NO 127 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 127 Ser Pro Asn Thr Asp Val Arg Met Tyr Ser Gly Lys Arg Asn Gly 5 10 15 <210> SEQ ID NO 128 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 128 Tyr Leu Ala Ala Pro Thr Gly Ile Pro Pro Ala Phe Phe Pro Ile 5 10 15 <210> SEQ ID NO 129 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 129 Gly Val Ala Ala Ala Thr Pro Arg Pro Asp Pro Glu Asp Gly Ala 5 10 15 <210> SEQ ID NO 130 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 130 Glu Glu Ile Pro Trp Val Arg Val Tyr Glu Asn Ile Cys Pro Arg 5 10 15 <210> SEQ ID NO 131 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 131 Pro Gly Asp Ala Pro Asp Ser Pro Tyr Ile Glu Ala Glu Asn Pro 5 10 15 <210> SEQ ID NO 132 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 132 Pro Asp Ser Pro Tyr Ile Glu Ala Glu Asn Pro Leu Tyr Asp Trp 5 10 15 <210> SEQ ID NO 133 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 133 Glu Asn Pro Leu Tyr Asp Trp Gly Gly Ser Ala Leu Phe Ser Pro 5 10 15 <210> SEQ ID NO 134 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 134 Ala Val Pro Leu Leu Ser Ala Gly Gly Ala Ala Pro Pro His Pro 5 10 15 <210> SEQ ID NO 135 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 135 Glu Leu Tyr Tyr Gly Pro Val Ser Pro Ala Asp Pro Glu Ser Pro 5 10 15 <210> SEQ ID NO 136 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 136 Pro Met Arg Ala Arg Pro Arg Gly Glu Val Arg Phe Leu His Tyr 5 10 15 <210> SEQ ID NO 137 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 137 Arg Pro Arg Gly Glu Val Arg Phe Leu His Tyr Asp Glu Ala Gly 5 10 15 <210> SEQ ID NO 138 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 138 Val Ala Gly Phe Asn Lys Arg Val Phe Cys Ala Ala Val Gly Arg 5 10 15 <210> SEQ ID NO 139 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 139 Ala Ile Asp Tyr Val His Cys Glu Gly Ile Ile His Arg Asp Ile 5 10 15 <210> SEQ ID NO 140 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 140 Ala Phe Pro Val Ala Leu His Ala Val Asp Ala Pro Ser Gln Phe 5 10 15 <210> SEQ ID NO 141 <211> LENGTH: 1808 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 141 atggaccggg aggcacttcg ggccatcagc cgcgggtgca agcccccttc gaccctggca 60 aaactggtga ccgggctggg attcgcgatc cacggagcgc tcatcccggg gtcggagggg 120 tgtgtctttg atagcagcca cccgaactac cctcatcggg taatcgtcaa ggcggggtgg 180 tacgccagca cgaaccacga ggcgcggctg ctgagacgcc tgaaccaccc cgcgatccta 240 cccctcctgg acctgcacgt cgtttctggg gtcacgtgtc tggtcctccc caagtatcac 300 tgcgacctgt atacctatct gagcaagcgc ccgtctccgt tgggccacct acagataacc 360 gcggtctccc ggcagctctt gagcgccatc gactacgtcc actgcgaagg catcatccac 420 cgcgatatta agaccgagaa catcctcatc aacacccccg agaacatctg tctgggggac 480 tttggggcgg cgtgctttgt gcgcgggtgt cgatcgagcc ccttccatta cgggatcgca 540 ggcaccatcg atacaaacgc ccccgaggtc ctggccgggg atccgtacac ccaggtaatc 600 gacatctgga gcgccggcct ggtgatcttt gagaccgccg tccacaccgc gtccttgttc 660 tcggccccgc gcgaccccga aaggcggccg tgcgacaacc agatcgcgcg catcatccga 720 caggcccagg tacacgtcga cgagtttcca acgcacgcgg aatcgcgcct caccgcgcac 780 taccgctcgc gggcggccgg gaacaatcgt ccggcgtgga cccgaccggc atggacccgc 840 tactacaaga tccacacaga cgtcgaatat ctcatctgca aagcccttac ctttgacgcg 900 gcgctccgcc caagcgccgc ggagttgctg cgcctgccgc tatttcaccc taagtgaccc 960 cgctcccccc ggggggcgtg gagggggggc tggttggatg tttttgcaca aaaagacgcg 1020 gccctcgggc tttggtgttt ttggcacctt gccgcccggc gtcatgcacg ccatcgctcc 1080 caggttgctt cttctttttg ttctttctgg tcttccgggg acacgcggcg ggtcgggtgt 1140 ccccggacca attaatcccc ccaacaacga tgttgttttc ccgggaggtt cccccgtggc 1200 tcaatattgt tatgcctatc cccggttgga cgatcccggg cccttgggtt ccgcggacgc 1260 cgggcggcaa gacctgcccc ggcgcgtcgt ccgtcacgag cccctgggcc gctcgttcct 1320 cacggggggg ctggttttgc tggcgccgcc ggtacgcgga tttggcgcac ccaacgcaac 1380 gtatgcggcc cgtgtgacgt actaccggct cacccgcgcc tgccgtcagc ccatcctcct 1440 tcggcagtat ggagggtgtc gcggcggcga gccgccgtcc ccaaagacgt gcgggtcgta 1500 cacgtacacg taccagggcg gcgggcctcc gacccggtac gctctcgtaa atgcttccct 1560 gctggtgccg atctgggacc gcgccgcgga gacattcgag taccagatcg aactcggcgg 1620 cgagctgcac gtgggtctgt tgtgggtaga ggtgggcggg gagggccccg gccccaccgc 1680 ccccccacag gcggcgcgtg cggagggcgg cccgtgcgtc cccccggtcc ccgcgggccg 1740 cccgtggcgc tcggtgcccc cggtatggta ttccgccccc aaccccgggt ttcgtggcct 1800 gcgtttcc 1808 <210> SEQ ID NO 142 <211> LENGTH: 248 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 142 Met His Ala Ile Ala Pro Arg Leu Leu Leu Leu Phe Val Leu Ser Gly 5 10 15 Leu Pro Gly Thr Arg Gly Gly Ser Gly Val Pro Gly Pro Ile Asn Pro 20 25 30 Pro Asn Asn Asp Val Val Phe Pro Gly Gly Ser Pro Val Ala Gln Tyr 35 40 45 Cys Tyr Ala Tyr Pro Arg Leu Asp Asp Pro Gly Pro Leu Gly Ser Ala 50 55 60 Asp Ala Gly Arg Gln Asp Leu Pro Arg Arg Val Val Arg His Glu Pro 65 70 75 80 Leu Gly Arg Ser Phe Leu Thr Gly Gly Leu Val Leu Leu Ala Pro Pro 85 90 95 Val Arg Gly Phe Gly Ala Pro Asn Ala Thr Tyr Ala Ala Arg Val Thr 100 105 110 Tyr Tyr Arg Leu Thr Arg Ala Cys Arg Gln Pro Ile Leu Leu Arg Gln 115 120 125 Tyr Gly Gly Cys Arg Gly Gly Glu Pro Pro Ser Pro Lys Thr Cys Gly 130 135 140 Ser Tyr Thr Tyr Thr Tyr Gln Gly Gly Gly Pro Pro Thr Arg Tyr Ala 145 150 155 160 Leu Val Asn Ala Ser Leu Leu Val Pro Ile Trp Asp Arg Ala Ala Glu 165 170 175 Thr Phe Glu Tyr Gln Ile Glu Leu Gly Gly Glu Leu His Val Gly Leu 180 185 190 Leu Trp Val Glu Val Gly Gly Glu Gly Pro Gly Pro Thr Ala Pro Pro 195 200 205 Gln Ala Ala Arg Ala Glu Gly Gly Pro Cys Val Pro Pro Val Pro Ala 210 215 220 Gly Arg Pro Trp Arg Ser Val Pro Pro Val Trp Tyr Ser Ala Pro Asn 225 230 235 240 Pro Gly Phe Arg Gly Leu Arg Phe 245 <210> SEQ ID NO 143 <211> LENGTH: 699 <212> TYPE: PRT <213> ORGANISM: HSV-2 <400> SEQUENCE: 143 Met His Ala Ile Ala Pro Arg Leu Leu Leu Leu Phe Val Leu Ser Gly 5 10 15 Leu Pro Gly Thr Arg Gly Gly Ser Gly Val Pro Gly Pro Ile Asn Pro 20 25 30 Pro Asn Ser Asp Val Val Phe Pro Gly Gly Ser Pro Val Ala Gln Tyr 35 40 45 Cys Tyr Ala Tyr Pro Arg Leu Asp Asp Pro Gly Pro Leu Gly Ser Ala 50 55 60 Asp Ala Gly Arg Gln Asp Leu Pro Arg Arg Val Val Arg His Glu Pro 65 70 75 80 Leu Gly Arg Ser Phe Leu Thr Gly Gly Leu Val Leu Leu Ala Pro Pro 85 90 95 Val Arg Gly Phe Gly Ala Pro Asn Ala Thr Tyr Ala Ala Arg Val Thr 100 105 110 Tyr Tyr Arg Leu Thr Arg Ala Cys Arg Gln Pro Ile Leu Leu Arg Gln 115 120 125 Tyr Gly Gly Cys Arg Gly Gly Glu Pro Pro Ser Pro Lys Thr Cys Gly 130 135 140 Ser Tyr Thr Tyr Thr Tyr Gln Gly Gly Gly Pro Pro Thr Arg Tyr Ala 145 150 155 160 Leu Val Asn Ala Ser Leu Leu Val Pro Ile Trp Asp Arg Ala Ala Glu 165 170 175 Thr Phe Glu Tyr Gln Ile Glu Leu Gly Gly Glu Leu His Val Gly Leu 180 185 190 Leu Trp Val Glu Val Gly Gly Glu Gly Pro Gly Pro Thr Ala Pro Pro 195 200 205 Gln Ala Ala Arg Ala Glu Gly Gly Pro Cys Val Pro Pro Val Pro Ala 210 215 220 Gly Arg Pro Trp Arg Ser Val Pro Pro Val Trp Tyr Ser Ala Pro Asn 225 230 235 240 Pro Gly Phe Arg Gly Leu Arg Phe Arg Glu Arg Cys Leu Pro Pro Gln 245 250 255 Thr Pro Ala Ala Pro Ser Asp Leu Pro Arg Val Ala Phe Ala Pro Gln 260 265 270 Ser Leu Leu Val Gly Ile Thr Gly Arg Thr Phe Ile Arg Met Ala Arg 275 280 285 Pro Thr Glu Asp Val Gly Val Leu Pro Pro His Trp Ala Pro Gly Ala 290 295 300 Leu Asp Asp Gly Pro Tyr Ala Pro Phe Pro Pro Arg Pro Arg Phe Arg 305 310 315 320 Arg Ala Leu Arg Thr Asp Pro Glu Gly Val Asp Pro Asp Val Arg Ala 325 330 335 Pro Arg Thr Gly Arg Arg Leu Met Ala Leu Thr Glu Asp Thr Ser Ser 340 345 350 Asp Ser Pro Thr Ser Ala Pro Glu Lys Thr Pro Leu Pro Val Ser Ala 355 360 365 Thr Ala Met Ala Pro Ser Val Asp Pro Ser Ala Glu Pro Thr Ala Pro 370 375 380 Ala Thr Thr Thr Pro Pro Asp Glu Met Ala Thr Gln Ala Ala Thr Val 385 390 395 400 Ala Val Thr Pro Glu Glu Thr Ala Val Ala Ser Pro Pro Ala Thr Ala 405 410 415 Ser Val Glu Ser Ser Pro Leu Pro Ala Ala Ala Ala Ala Thr Pro Gly 420 425 430 Ala Gly His Thr Asn Thr Ser Ser Ala Ser Ala Ala Lys Thr Pro Pro 435 440 445 Thr Thr Pro Ala Pro Thr Thr Pro Pro Pro Thr Ser Thr His Ala Thr 450 455 460 Pro Arg Pro Thr Thr Pro Gly Pro Gln Thr Thr Pro Pro Gly Pro Ala 465 470 475 480 Thr Pro Gly Pro Val Gly Ala Ser Ala Ala Pro Thr Ala Asp Ser Pro 485 490 495 Leu Thr Ala Ser Pro Pro Ala Thr Ala Pro Gly Pro Ser Ala Ala Asn 500 505 510 Val Ser Val Ala Ala Thr Thr Ala Thr Pro Gly Thr Arg Gly Thr Ala 515 520 525 Arg Thr Pro Pro Thr Asp Pro Lys Thr His Pro His Gly Pro Ala Asp 530 535 540 Ala Pro Pro Gly Ser Pro Ala Pro Pro Pro Pro Glu His Arg Gly Gly 545 550 555 560 Pro Glu Glu Phe Glu Gly Ala Gly Asp Gly Glu Pro Pro Glu Asp Asp 565 570 575 Asp Ser Ala Thr Gly Leu Ala Phe Arg Thr Pro Asn Pro Asn Lys Pro 580 585 590 Pro Pro Ala Arg Pro Gly Pro Ile Arg Pro Thr Leu Pro Pro Gly Ile 595 600 605 Leu Gly Pro Leu Ala Pro Asn Thr Pro Arg Pro Pro Ala Gln Ala Pro 610 615 620 Ala Lys Asp Met Pro Ser Gly Pro Thr Pro Gln His Ile Pro Leu Phe 625 630 635 640 Trp Phe Leu Thr Ala Ser Pro Ala Leu Asp Ile Leu Phe Ile Ile Ser 645 650 655 Thr Thr Ile His Thr Ala Ala Phe Val Cys Leu Val Ala Leu Ala Ala 660 665 670 Gln Leu Trp Arg Gly Arg Ala Gly Arg Arg Arg Tyr Ala His Pro Ser 675 680 685 Val Arg Tyr Val Cys Leu Pro Pro Glu Arg Asp 690 695 <210> SEQ ID NO 144 <211> LENGTH: 1599 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 144 atggagctca gctatgccac caccctgcac caccgggacg ttgtgtttta cgtcacggca 60 gacagaaacc gcgcctactt tgtgtgcggg gggtccgttt attccgtagg gcggcctcgg 120 gattctcagc cgggggaaat tgccaagttt ggcctggtgg tccgggggac aggccccaaa 180 gaccgcatgg tcgccaacta cgtacgaagc gagctccgcc agcgcggcct gcgggacgtg 240 cggcccgtgg gggaggacga ggtgttcctg gacagcgtgt gtctgctaaa cccgaacgtg 300 agctccgagc gagacgtgat taataccaac gacgttgaag tgctggacga atgcctggcc 360 gaatactgca cctcgctgcg aaccagcccg ggggtgctgg tgaccggggt gcgcgtgcgc 420 gcgcgagaca gggtcatcga gctatttgag cacccggcga tcgtcaacat ttcctcgcgc 480 ttcgcgtaca ccccctcccc ctacgtattc gccctggccc aggcgcacct cccccggctc 540 ccgagctcgc tggagcccct ggtgagcggc ctgtttgacg gcattcccgc cccgcgccag 600 cccctggacg cccgcgaccg gcgcacggat gtcgtgatca cgggcacccg cgcccccaga 660 ccgatggccg ggaccggggc cgggggcgcg ggggccaagc gggccaccgt cagcgagttc 720 gtgcaagtga agcacatcga ccgtgttgtg tccccgagcg tctcttccgc ccccccgccg 780 agcgcccccg acgcgagtct gccgcccccg gggctccagg aggccgcccc gccgggcccc 840 ccgctcaggg agctgtggtg ggtgttctac gccggcgacc gggcgctgga ggagccccac 900 gccgagtcgg gattgacgcg cgaggaggtc cgcgccgtgc atgggttccg ggagcaggcg 960 tggaagctgt ttgggtcggt gggggctccg cgggcgtttc tcggggccgc gctggccctg 1020 agcccgaccc aaaagctcgc cgtctactac tatctcatcc accgggagcg gcgcatgtcc 1080 cccttccccg cgctcgtgcg gctcgtcggt cggtacatcc agcgccacgg cctgtacgtt 1140 cccgcgcccg acgaaccgac gttggccgat gccatgaacg ggctgttccg cgacgcgctg 1200 gcggccggga ccgtggccga gcagctcctc atgttcgacc tcctcccgcc caaggacgtg 1260 ccggtgggga gcgacgcgcg ggccgacagc gccgccctgc tgcgctttgt ggactcgcaa 1320 cgcctgaccc cgggggggtc cgtctcgccc gagcacgtca tgtacctcgg cgcgttcctg 1380 ggcgtgttgt acgccggcca cggacgcctg gccgcggcca cgcataccgc gcgcctgacg 1440 ggcgtgacgt ccctggtcct gaccgtgggg gacgtcgacc ggatgtccgc gtttgaccgc 1500 gggccggcgg gggcggctgg ccgcacgcga accgccgggt acctggacgc gctgcttacc 1560 gtttgcctgg ctcgcgccca gcacggccag tctgtgtga 1599 <210> SEQ ID NO 145 <211> LENGTH: 1110 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 145 atgagtcagt gggggcccag ggcgatcctt gtccagacgg acagcaccaa ccggaatgcc 60 gatggggact ggcaagcggc cgtagctatt cgcgggggcg gagtcgttca actgaacatg 120 gtcaacaaac gcgccgtgga ttttaccccg gcagaatgcg gggactccga atgggccgtg 180 ggccgcgtct ctctgggcct gcgaatggca atgccgcggg acttctgcgc gattattcac 240 gcccccgcgg tatccggccc cgggccccac gtgatgctcg gtctcgtcga ctcgggctac 300 cgcggaaccg tcctggccgt ggtcgtagcc ccgaacggga cgcgcgggtt tgcccccggg 360 gccctccggg tcgacgtgac gtttctggac atccgggcca cccccccgac cctcaccgag 420 ccgagctccc tgcaccggtt tccgcagttg gcgccgtccc cgctggcagg gttacgagaa 480 gatccttggt tggacggggc gctcgcgacc gccggggggg cggtggccct gccggccaga 540 cggcgcgggg gatcgctggt ctacgcgggc gagctaacgc aggtgaccac cgagcacggc 600 gactgcgtgc acgaggcgcc cgcctttctg ccaaagcgcg aggaggacgc aggctttgac 660 attctcatcc accgagccgt gaccgtcccg gccaacggcg ccacggtcat acagccgtcc 720 ctccgcgtat tgcgcgcggc cgacggacca gaggcctgct atgtgctggg gcggtcgtcg 780 ctcaatgcca ggggcctcct ggtcatgcct acgcgctggc cctccgggca cgcctgtgcg 840 tttgttgtat gtaacctgac cggagtcccg gtgaccctac aagccgggtc caaggtcgcc 900 cagctgctcg tcgcggggac ccacgccctc ccctggatcc cccccgacaa catccacgag 960 gacggcgcat tccgggccta ccccagaggg gttccggacg cgaccgccac cccccgagac 1020 ccgccgattt tggtgtttac gaacgagttt gacgcggacg cccccccaag caagcggggg 1080 gccggggggt ttggctccac tggcatctag 1110 <210> SEQ ID NO 146 <211> LENGTH: 1446 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 146 atggcctgtc gtaagttctg tggggtctac cgtagacccg acaagagaca ggaggcgtcc 60 gtcccgccgg agacaaacac ggccccggcc ttcccggcga gcacctttta tacccccgcg 120 gaggatgcgt acctggcccc cgggcccccg gaaaccatcc acccttcccg cccaccgtcc 180 cccggcgagg ctgcgcgcct gtgtcagctg caggagatct tggcccagat gcacagcgac 240 gaggactacc ccatcgtgga cgccgcgggt gcggaggagg aagacgaggc cgacgatgac 300 gccccggatg acgtggccta cccggaggac tacgcggagg ggcgttttct gtccatggtt 360 tcggccgccc ccctgcccgg agccagcggc catcctcctg ttccgggccg cgcagccccc 420 cccgacgtcc ggacctgcga cacgggtaag gtgggggcca cggggttcac cccggaagag 480 ctcgacacca tggaccggga ggcacttcgg gccatcagcc gcgggtgcaa gcccccttcg 540 accctggcaa aactggtgac cgggctggga ttcgcgatcc acggagcgct catcccgggg 600 tcggaggggt gtgtctttga tagcagccac ccgaactacc ctcatcgggt aatcgtcaag 660 gcggggtggt acgccagcac gagccacgag gcgcggctgc tgagacgcct gaaccacccc 720 gcgatcctac ccctcctgga cctgcacgtc gtttctgggg tcacgtgtct ggtcctcccc 780 aagtatcact gcgacctgta tacctatctg agcaagcgcc cgtctccgtt gggccaccta 840 cagataaccg cggtctcccg gcagctcttg agcgccatcg actacgtcca ctgcaaaggc 900 atcatccacc gcgatattaa gaccgagaac atcttcatca acacccccga gaacatctgt 960 ctgggggact ttggggcggc gtgctttgtg cgcgggtgtc gatcgagccc cttccattac 1020 gggatcgcag gcaccatcga tacaaacgcc cccgaggtcc tggccgggga tccgtacacc 1080 caggtaatcg acatctggag cgccggcctg gtgatctttg agaccgccgt ccacaccgcg 1140 tccttgttct cggccccgcg cgaccccgaa aggcggccgt gcgacaacca gatcgcgcgc 1200 atcatccgac aggcccaggt acacgtcgac gagtttccga cgcacgcgga atcgcgcctc 1260 accgcgcact accgctcgcg ggcggccggg aacaatcgtc cggcgtggac ccgaccggcg 1320 tggacccgct actacaagat ccacacagac gtcgaatatc tcatatgcaa agcccttacc 1380 tttgacgcgg cgctccgccc aagcgccgcg gagttgctgc gcctgccgct atttcaccct 1440 aagtga 1446 <210> SEQ ID NO 147 <211> LENGTH: 1539 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 147 atggctaccg acattgatat gctaatcgac ctaggattgg acctgtccga cagcgagctc 60 gaggaggacg ctctggagcg ggacgaggag ggccgccgcg acgaccccga gtccgacagc 120 agcggggagt gttcctcgtc ggacgaggac atggaagacc cctgcggaga cggaggggcg 180 gaggccatcg acgcggcgat tcccaaaggt cccccggccc gccccgagga cgccggcacc 240 cccgaagcct cgacgcctcg cccggcagcg cggcggggag ccgacgatcc gccacccgcg 300 accaccggcg tgtggtcgcg cctcgggacc aggcggtcgg cttccccccg ggaaccgcac 360 ggggggaagg tggcccgcat ccaacccccg tcgaccaagg caccgcatcc ccgaggcggg 420 cggcgaggtc gccgccgggg ccggggtcga tacggccccg gcggcgccga ctccacacca 480 aaaccccgcc ggcgcgtctc cagaaacgcc cacaaccaag ggggtcgcca ccccgcgtcg 540 gcgcggacgg acggccccgg cgccacccac ggcgaggcgc ggcgcggagg ggagcagctc 600 gacgtctccg ggggcccgcg gccacgaggc acgcgccagg ccccccctcc gctgatggcg 660 ctgtccctga cccccccgca cgcggacggc cgcgccccgg tcccggagcg aaaggcgccc 720 tctgccgaca ccatcgaccc cgccgttcgg gcggttctgc gatccatatc cgagcgcgcg 780 gcggtcgagc gcatcagcga aagctttgga cgcagtgccc tggtcatgca agaccccttt 840 ggcgggatgc cgtttcccgc cgcgaacagc ccctgggctc ccgtgctggc cacccaagcg 900 ggggggtttg acgccgagac ccgtcgggtt tcctgggaaa ccctggtcgc tcacggcccg 960 agcctctacc gcacattcgc agccaacccg cgggccgcgt cgacagccaa ggccatgcgc 1020 gactgcgtgc tgcgccagga aaatctcatc gaggccctgg cgtccgcgga tgagacgctg 1080 gcgtggtgca agatgtgcat tcaccacaat ctgccgctcc gcccccagga ccctatcatc 1140 ggaacggcgg ccgccgtgct ggaaaacctc gccacgcgcc tgcgcccctt tctgcagtgc 1200 tacctgaagg cccgaggcct gtgcgggctg gacgacctgt gctcgcggcg acgcctgtcg 1260 gacattaagg atattgcctc ctttgtgttg gtcatcctgg cccgcctcgc caaccgcgtc 1320 gagcgcggcg tgtcggagat cgactacacg accgtggggg ttggggccgg cgagacgatg 1380 cacttttaca tcccgggggc ctgcatggcg ggtctcattg aaatactgga cacgcaccgc 1440 caggagtgtt ccagtcgcgt gtgcgagctg acggccagtc acactatcgc ccccttatat 1500 gtgcacggca aatacttcta ctgcaactcc ctattttag 1539 <210> SEQ ID NO 148 <211> LENGTH: 1638 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 148 atggctcgcg gggccgggtt ggtgtttttt gttggagttt gggtcgtatc gtgcctggcg 60 gcagcaccca gaacgtcctg gaaacgggta acctcgggcg aggacgtggt gttgcttccg 120 gcgcccgcgg aacgcacccg ggcccacaaa ctactgtggg ccgcggaacc cctggatgcc 180 tgcggtcccc tgcgcccgtc gtgggtggcg ctgtggcccc cccgacgggt gctcgagacg 240 gtcgtggatg cggcgtgcat gcgcgccccg gaaccgctcg ccatagcata cagtcccccg 300 ttccccgcgg gcgacgaggg actgtattcg gagttggcgt ggcgcgatcg cgtagccgtg 360 gtcaacgaga gtctggtcat ctacggggcc ctggagacgg acagcggtct gtacaccctg 420 tccgtggtcg gcctaagcga cgaggcgcgc caagtggcgt cggtggttct ggtcgtggag 480 cccgcccctg tgccgacccc gacccccgac gactacgacg aagaagacga cgcgggcgtg 540 acgaacgcac gccggtcagc gttccccccc caaccccccc cccgtcgtcc ccccgtcgcc 600 cccccgacgc accctcgtgt tatccccgag gtgtcccacg tgcgcggggt aacggtccat 660 atggagaccc tggaggccat tctgtttgcc cccggggaga cgtttgggac gaacgtctcc 720 atccacgcca ttgcccacga cgacggtccg tacgccatgg acgtcgtctg gatgcggttt 780 gacgtgccgt cctcgtgcgc cgatatgcgg atctacgaag cttgtctgta tcacccgcag 840 cttccagagt gtctatctcc ggccgacgcg ccgtgcgccg taagttcctg ggcgtaccgc 900 ctggcggtcc gcagctacgc cggctgttcc aggactacgc ccccgccgcg atgttttgcc 960 gaggctcgca tggaaccggt cccggggttg gcgtggctgg cctccaccgt caatctggaa 1020 ttccagcacg cctcccccca gcacgccggc ctctacctgt gcgtggtgta cgtggacgat 1080 catatccacg cctggggcca catgaccatc agcaccgcgg cgcagtaccg gaacgcggtg 1140 gtggaacagc acctccccca gcgccagccc gagcccgtcg agcccacccg cccgcacgtg 1200 agagcccccc atcccgcgcc ctccgcgcgc ggcccgctgc gcctcggggc ggtgctgggg 1260 gcggccctgt tgctggccgc cctcgggctg tccgcgtggg cgtgcatgac ctgctggcgc 1320 aggcgctcct ggcgggcggt taaaagccgg gcctcggcga cgggccccac ttacattcgc 1380 gtggcggaca gcgagctgta cgcggactgg agttcggaca gcgaggggga gcgcgacggg 1440 tccctgtggc aggaccctcc ggagagaccc gactctccct ccacaaatgg atccggcttt 1500 gagatcttat caccaacggc tccgtctgta tacccccata gcgaggggcg taaatctcgc 1560 cgcccgctca ccacctttgg ttcgggaagc ccgggccgtc gtcactccca ggcctcctat 1620 ccgtccgtcc tctggtaa 1638 <210> SEQ ID NO 149 <211> LENGTH: 4125 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 149 atggccgctc ctgcccgcga ccccccgggt taccggtacg ccgcggccat cctgcccacc 60 ggctccatcc tgagtacgat cgaggtggcg tcccaccgca gactctttga ttttttcgcc 120 gccgtgcgct ccgacgaaaa cagcctgtat gacgtagagt ttgacgccct gctggggtcc 180 tactgcaaca ccctgtcgct cgtgcgcttt ctggagctcg gcctgtccgt ggcgtgcgtg 240 tgcaccaagt tcccggagct ggcttacatg aacgaagggc gtgtgcagtt cgaggtccac 300 cagcccctca tcgcccgcga cggcccgcac cccgtcgagc agcccgtgca taattacatg 360 acgaaggtca tcgaccgccg ggccctgaac gccgccttca gcctggccac cgaggccatt 420 gccctgctca cgggggaggc cctggacggg acgggcatta gcctgcatcg ccagctgcgc 480 gccatccagc agctcgcgcg caacgtccag gccgtcctgg gggcgtttga gcgcggcacg 540 gccgaccaga tgctgcacgt gctgttggag aaggcgcctc ccctggccct gctgttgccc 600 atgcaacgat atctcgacaa cgggcgcctg gcgaccaggg ttgcccgggc gaccctggtc 660 gccgagctga agcggagctt ttgcgacacg agcttcttcc tgggcaaggc gggccatcgc 720 cgcgaggcca tcgaggcctg gctcgtggac ctgaccacgg cgacgcagcc gtccgtggcc 780 gtgccccgcc tgacgcacgc cgacacgcgc gggcggccgg tcgacggggt gctggtcacc 840 accgccgcca tcaaacagcg cctcctgcag tccttcctga aggtggagga caccgaggcc 900 gacgtgccgg tgacctacgg cgagatggtc ttgaacgggg ccaacctcgt cacggcgctg 960 gtgatgggca aggccgtgcg gagcctggac gacgtgggcc gccacctgct ggatatgcag 1020 gaggagcaac tcgaggcgaa ccgggagacg ctggatgaac tcgaaagcgc cccccagaca 1080 acgcgcgtgc gcgcggatct ggtggccata ggcgacaggc tggtcttcct ggaggccctg 1140 gagagacgca tctacgccgc caccaacgtg ccctaccccc tggtgggcgc catggacctg 1200 acgttcgtcc tgcccctggg gctgttcaac ccggccatgg agcgcttcgc cgcgcacgcc 1260 ggggacctgg tgcccgcccc cggccacccg gagccccgcg cgttccctcc ccggcagctg 1320 tttttttggg gaaaggacca ccaggttctg cggctgtcca tggagaacgc ggtcgggacc 1380 gtgtgtcatc cttcgctcat gaacatcgac gcggccgtcg ggggcgtgaa ccacgacccc 1440 gtcgaggccg cgaatccgta cggggcgtac gtcgcggccc cggccggccc cggcgcggac 1500 atgcagcagc gttttctgaa cgcctggcgg cagcgcctcg cccacggccg ggtccggtgg 1560 gtcgccgagt gccagatgac cgcggagcag ttcatgcagc ccgacaacgc caacctggct 1620 ctggagctgc accccgcgtt cgacttcttc gcgggcgtgg ccgacgtcga gcttcccggc 1680 ggcgaagtcc ccccggccgg tccgggggcg atccaggcca cctggcgcgt ggtcaacggc 1740 aacctgcccc tggcgctgtg tccggtggcg tttcgtgacg cccggggcct ggagctcggc 1800 gttggccgcc acgccatggc gccggctacc atagccgccg tccgcggggc gttcgaggac 1860 cgcagctacc cggcggtgtt ttacctgctg caagccgcga ttcacggcaa cgagcacgtg 1920 ttctgcgccc tggcgcggct cgtgactcag tgcatcacca gctactggaa caacacgcga 1980 tgcgcggcgt tcgtgaacga ctactcgctg gtctcgtaca tcgtgaccta cctcgggggc 2040 gacctccccg aggagtgcat ggccgtgtat cgggacctgg tggcccacgt cgaggccctg 2100 gcccagctgg tggacgactt taccctgccg ggcccggagc tgggcgggca ggctcaggcc 2160 gagctgaatc acctgatgcg cgacccggcg ctgctgccgc ccctcgtgtg ggactgcgac 2220 ggccttatgc gacacgcggc cctggaccgc caccgagact gccggattga cgcggggggg 2280 cacgagcccg tctacgcggc ggcgtgcaac gtggcgacgg ccgactttaa ccgcaacgac 2340 ggccggctgc tgcacaacac ccaggcccgc gcggccgacg ccgccgacga ccggccgcac 2400 cggccggccg actggaccgt ccaccacaaa atctactatt acgtgctggt gccggccttc 2460 tcgcgggggc gctgctgcac cgcgggggtc cgcttcgacc gcgtgtacgc cacgctgcag 2520 aacatggtgg tcccggagat cgcccccggt gaggagtgcc cgagcgatcc cgtgaccgac 2580 cccgcccacc cgctgcatcc cgccaatctg gtggccaaca cggtcaagcg catgttccac 2640 aacgggcgcg tcgtcgtcga cgggcccgcc atgctcacgc tgcaggtgct ggcgcacaac 2700 atggccgagc gcacgacggc gctgctgtgc tccgcggcgc ccgacgcggg cgccaacacc 2760 gcgtcgacgg ccaacatgcg catcttcgac ggggcgctgc acgccggcgt gctgctcatg 2820 gccccccagc acctggacca caccatccaa aatggcgaat acttctacgt cctgcccgtc 2880 cacgcgctgt ttgcgggcgc cgaccacgtg gccaacgcgc ccaacttccc cccggccctg 2940 cgcgacctgg cgcgcgacgt ccccctggtc cccccggccc tgggggccaa ctacttctcc 3000 tccatccgcc agcccgtggt gcagcacgcc cgcgagagcg cggcggggga gaacgcgctg 3060 acctacgcgc tcatggcggg gtacttcaag atgagccccg tggccctgta tcaccagctc 3120 aagacgggcc tccaccccgg gttcgggttc accgtcgtgc ggcaggaccg cttcgtgacc 3180 gagaacgtgc tgttttccga gcgcgcgtcg gaggcgtact ttctgggcca gctccaggtg 3240 gcccgccacg aaacgggcgg gggggtcaac ttcacgctca cccagccgcg cggaaacgtg 3300 gacctgggtg tgggctacac cgccgtcgcg gccacgggca ccgtccgcaa ccccgttacg 3360 gacatgggca acctccccca aaacttttac ctcggccgcg gggccccccc gctgctagac 3420 aacgcggccg ccgtgtacct gcgcaacgcg gtcgtggcgg gaaaccggct ggggccggcc 3480 cagcccctcc cggtctttgg ctgcgcccag gtgccgcggc gcgccggcat ggaccacggg 3540 caggatgccg tgtgtgagtt catcgccacc cccgtggcca cggacatcaa ctactttcgc 3600 cggccctgca acccgcgggg acgcgcggcc ggcggcgtgt acgcggggga caaggagggg 3660 gacgtcatag ccctcatgta cgaccacggc cagagcgacc cggcgcggcc cttcgcggcc 3720 acggccaacc cgtgggcgtc gcagcggttc tcgtacgggg acctgctgta caacggggcc 3780 tatcacctca acggggcctc gcccgtcctc agcccctgct tcaagttctt caccgcggcc 3840 gacatcacgg ccaaacatcg ctgcctggag cgtctcatcg tggaaacggg atcggcggta 3900 tccacggcca ccgctgccag cgacgtgcag tttaagcgcc cgccggggtg ccgcgagctc 3960 gtggaagacc cgtgcggcct gtttcaggaa gcctacccga tcacctgcgc cagcgacccc 4020 gccctgctac gcagcgcccg cgatggggag gcccacgcgc gagagaccca ctttacgcag 4080 tatctcatct acgacgcctc cccgctaaag ggcctgtctc tgtaa 4125 <210> SEQ ID NO 150 <211> LENGTH: 2169 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 150 atgcaacgcc gggcgcgcgg cgcgagctcc ctgcggctgg cgcggtgcct gacgccggcc 60 aacctcatcc gcggcgccaa cgcgggcgtc cccgagcggc gcatcttcgc cgggtgtctg 120 ctccccaccc cggaggggct cctcagcgcg gccgtgggcg tcctgcggca gcgcgccgac 180 gacctgcagc cggcgtttct gaccggcgcc gatcgcagcg tccggctggc ggcgcggcac 240 cataacaccg tccccgagag cctgatcgta gacgggctcg ccagcgaccc gcactacgac 300 tacatccggc actacgcgtc ggccgccaag caggcgctcg gcgaggtgga gctgtcgggc 360 ggccagctga gccgcgccat cctagcgcag tactggaagt acctccagac ggtcgtgccc 420 agcggcctgg acatccccga cgacccggcg ggcgactgcg accccagcct gcacgtgctg 480 ctgcggccca ccctgctccc gaagctgctg gtgcgcgccc cgttcaagag cggggccgcc 540 gcggccaagt acgccgccgc ggtggcgggg ttgcgcgacg cggcccacag gctccagcag 600 tacatgttct ttatgcgccc cgcagacccg agccggccga gcacggacac cgcactgcgg 660 ctgagcgagc tcctggccta cgtctccgtg ttgtaccatt gggcctcgtg gatgctgtgg 720 acggcggaca agtacgtgtg tcgccgcctg ggccccgccg atcgccggtt cgtggcgctc 780 agcgggagtc tggaggcgcc cgcggagacg tttgcgcgcc acctggaccg cgggcccagc 840 ggcaccacgg gctcgatgca gtgcatggcc ctgcgggcgg cggtcagcga cgtcctgggc 900 cacctgacgc gcctggccca cctgtgggag accggcaagc gcagcggcgg cacgtacggg 960 atcgtggacg ccatcgtctc gaccgtcgag gttctatcca tagtccacca ccacgcccag 1020 tatataatta acgcgacgct taccgggtat gtcgtctggg cctccgacag cctgaacaac 1080 gagtacctta cggcggcggt ggacagccag gagcgcttct gcaggaccgc cgcccccctg 1140 ttccccacga tgaccgcccc gagctgggcc cggatggaac tcagcatcaa gtcctggttc 1200 ggggccgccc tggccccgga cctgcttcgg agcggaaccc cgtcgcccca ctacgagtcc 1260 atcctgcgcc tcgcggcgtc cggcccaccg gggggccgcg gcgcggtcgg cgggagctgc 1320 cgggacaaga tacaacggac ccggcgcgac aacgcacccc cgccgctccc ccgggctcgc 1380 ccccactcga cccccgcggc ccctcggagg tgcaggcgcc accgcgagga cctccccgag 1440 cccccgcacg tcgacgcggc cgaccggggt cccgagccct gcgccggccg gccggccacg 1500 tattacacgc atatggccgg ggcgcccccg cgcctcccgc cccgcaaccc cgcgcccccc 1560 gagcagcggc cggcagccgc ggcgcgcccg ctcgcggctc agcgcgaggc cgccggggtc 1620 tacgacgcgg tgcggacctg ggggccggac gcggaggccg aaccggacca gatggaaaac 1680 acgtatctgc tgcccgacga tgacgccgcc atgcccgcgg gcgtcgggct tggcgccacc 1740 cccgccgccg acaccaccgc cgccgccgcc tggccggccg aaagccacgc cccccgcgcc 1800 ccctccgagg acgcagattc catttacgag tcggtgggcg aggatggggg gcgcgtctac 1860 gaggagatcc cctgggttcg ggtatacgaa aacatctgcc ctcgccggcg tcttgccggc 1920 ggggccgccc tgccgggaga cgccccggac tccccgtaca tcgaggcgga aaatcccctg 1980 tacgactggg gcgggtctgc cctcttctcc cctcggcggg ccacacgcgc cccggacccg 2040 ggactaagcc tgtcgcccat gcccgcccgc ccccggacca acgcgctggc caacgacggc 2100 ccgacgaacg tcgccgccct cagcgccctg ttgacgaagc tcaaacgcgg ccgacaccag 2160 agccattaa 2169 <210> SEQ ID NO 151 <211> LENGTH: 957 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 151 atgatcacgg attgtttcga agcagacatc gcgatcccct cgggtatctc gcgccccgat 60 gccgcggcgc tgcagcggtg cgagggtcga gtggtctttc tgccgaccat ccgccgccag 120 ctggcgctcg cggacgtggc gcacgaatcg ttcgtctccg gaggagttag tcccgacacg 180 ttggggttgt tgctggcgta ccgcaggcgc ttccccgcgg taatcacgcg ggtgctgccc 240 acgcgaatcg tcgcctgccc cgtggacctg gggctcacgc acgccggcac cgtcaatctc 300 cgcaacacct cccccgtcga cctctgcaac ggggatcccg tcagcctcgt cccgcccgtc 360 ttcgagggcc aggcgacgga cgtgcgcctg gagtcgctgg acctcacgct gcggtttccg 420 gtcccgctcc caacgcccct ggcccgcgag atagtcgcgc ggctggtcgc ccggggcatc 480 cgggacctca accccgaccc ccggacgccc ggggagctcc ccgacctcaa cgtgctgtat 540 tacaacgggg cccgtctctc gctcgtggcc gacgtccagc aactcgcctc cgtaaacacc 600 gagctgcggt cgctcgtcct caacatggtc tactccataa ccgaaggaac caccctcatc 660 ctcacgctca tcccccggct gctcgcgctg agcgcccagg acggatacgt gaacgcgctc 720 ctgcagatgc agagcgtcac gcgagaagcc gcccagctca tccaccccga agcccccatg 780 ctgatgcagg acggcgaacg caggctgccg ctttacgagg cgctggtcgc ctggctggcg 840 cacgcgggcc aactcgggga catcctggcc ctggccccgg cggttcgggt gtgtacgttc 900 gacggcgccg ccgttgtgca atccggcgac atggccccgg ttatccgcta cccctga 957 <210> SEQ ID NO 152 <211> LENGTH: 3066 <212> TYPE: DNA <213> ORGANISM: HSV2 <400> SEQUENCE: 152 atggaacccc ggcccggcac gagctcccgg gcggaccccg gccccgagcg gccgccgcgg 60 cagacccccg gcacggtgag agggcgaccc ccgggtctca ggccccccct tttccccgga 120 ccacccggct gcgggttggg ggtggtcgcg ggcggtgggc tcgggggcgg ggacgcttga 180 cggggccgac ccccggcccg cttaagcggt cgggggaccc ccgtgggccg tgcgccgccc 240 cccgaccctc tgggggggcg agggaggcag ggaggagccc gagagcgggg gacagggggg 300 gagacgaggg gtcggaatcc aaaggacgca gaccaccttt ggttacggac ccctttctcc 360 cccccttccg aacaaaaagc agcgggcggg gggccggggt gagggaggga cacgggggac 420 acggcgcggg ggtcccgcct cacgccccgc gccctctaaa tcccccccgt tgctttgtca 480 agcagcccgc cgccccgcac gcctggggga tgctcaacga catgcagtgg ctcgccagca 540 gcgactcgga ggaggagacc gaggtgggaa tctctgacga cgaccttcac cgcgactcca 600 cctccgaggc gggcagcacg gacacggaga tgttcgaggc gggcctgatg gacgcggcca 660 cgcccccggc ccggcccccg gccgagcgcc agggcagccc cacgcccgcc gacgcgcagg 720 gatcctgtgg gggtgggccc gtgggtgagg aggaagcgga agcgggaggg gggggcgacg 780 tgtgtgccgt gtgcacggac gagatcgccc cgcccctgcg ctgccagagt tttccctgcc 840 tgcacccctt ctgcatcccg tgcatgaaga cctggattcc gttgcgcaac acgtgtcccc 900 tgtgcaacac cccggtggcg tacctgatag tgggcgtgac cgccagcggg tcgttcagca 960 ccatcccgat agtgaacgac ccccggaccc gcgtggaggc cgaggcggcc gtgcgggccg 1020 gcacggccgt ggactttatc tggacgggca acccgcggac ggccccgcgc tccctgtcgc 1080 tggggggaca cacggtccgc gccctgtcgc ccaccccccc gtggcccggc acggacgacg 1140 aggacgatga cctggccgac ggtgagggcg ggcgggggtc gggcgggggg cgggcggggg 1200 tcgggcgggg gtcgggcggg ggtcgggcgg gggtcgggcg ggggtcgggc gggggtcggg 1260 cgggggtcgg gcgggggtcg ggcgggggtc gggcgggggt cgggcactaa ccgggggctc 1320 ccgtctctgt ctccctctgc agtggactac gtcccgcccg ccccccgaag agcgccccgg 1380 cgcgggggcg gcggtgcggg ggcgacccgc ggaacctccc agcccgccgc gacccgaccg 1440 gcgccccctg gcgccccgcg gagcagcagc agcggcggcg ccccgttgcg ggcgggggtg 1500 ggatctgggt ctgggggcgg ccctgccgtc gcggccgtcg tgccgagagt ggcctctctt 1560 ccccctgcgg ccggcggggg gcgcgcgcag gcgcggcggg tgggcgaaga cgccgcggcg 1620 gcggagggca ggacgccccc cgcgagacag ccccgcgcgg cccaggagcc ccccatagtc 1680 atcagcgact ctcccccgcc gtctccgcgc cgccccgcgg gccccgggcc gctctccttt 1740 gtctcctcct cctccgcaca ggtgtcctcg ggccccgggg ggggaggtct gccacagtcg 1800 tcggggcgcg ccgcgcgccc ccgcgcggcc gtcgccccgc gcgtccggag tccgccccgc 1860 gccgccgccg cccccgtggt gtctgcgagc gcggacgcgg ccgggcccgc gccgcccgcc 1920 gtgccggtgg acgcgcaccg cgcgccccgg tcgcgcatga cccaggctca gaccgacacc 1980 caagcacaga gtctgggccg ggcaggcgcg accgacgcgc gcgggtcggg agggccgggc 2040 gcggagggag gacccggggt cccccgcggc accaacaccc ccggtgccgc cccccacgcc 2100 gcggaggggg cggcggcccg cccccggaag aggcgcgggt cggactcggg ccccgcggcc 2160 tcgtcctccg cctcttcctc cgccgccccg cgctcgcccc tcgcccccca gggggtgggg 2220 gccaagaggg cggcgccgcg ccgggccccg gactcggact cgggcgaccg cggccacggg 2280 ccgctcgccc cggcgtccgc gggcgccgcg cccccgtcgg cgtctccgtc gtcccaggcc 2340 gcggtcgccg ccgcctcctc ctcctccgcc tcctcctcct ccgcctcctc ctcctccgcc 2400 tcctcctcct ccgcctcctc ctcctccgcc tcctcctcct ccgcctcctc ctcctccgcc 2460 tcttcctctg cgggcggggc tggtgggagc gtcgcgtccg cgtccggcgc tggggagaga 2520 cgagaaacct ccctcggccc ccgcgctgct gcgccgcggg ggccgaggaa gtgtgccagg 2580 aagacgcgcc acgcggaggg cggccccgag cccggggccc gcgacccggc gcccggcctc 2640 acgcgctacc tgcccatcgc gggggtctcg agcgtcgtgg ccctggcgcc ttacgtgaac 2700 aagacggtca cgggggactg cctgcccgtc ctggacatgg agacgggcca cataggggcc 2760 tacgtggtcc tcgtggacca gacggggaac gtggcggacc tgctgcgggc cgcggccccc 2820 gcgtggagcc gccgcaccct gctccccgag cacgcgcgca actgcgtgag gccccccgac 2880 tacccgacgc cccccgcgtc ggagtggaac agcctctgga tgaccccggt gggcaacatg 2940 ctctttgacc agggcaccct ggtgggcgcg ctggacttcc acggcctccg gtcgcgccac 3000 ccgtggtctc gggagcaggg cgcgcccgcg ccggccggcg acgcccccgc gggccacggg 3060 gagtag 3066 <210> SEQ ID NO 153 <211> LENGTH: 369 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 153 Met Ser Gln Trp Gly Pro Arg Ala Ile Leu Val Gln Thr Asp Ser Thr 5 10 15 Asn Arg Asn Ala Asp Gly Asp Trp Gln Ala Ala Val Ala Ile Arg Gly 20 25 30 Gly Gly Val Val Gln Leu Asn Met Val Asn Lys Arg Ala Val Asp Phe 35 40 45 Thr Pro Ala Glu Cys Gly Asp Ser Glu Trp Ala Val Gly Arg Val Ser 50 55 60 Leu Gly Leu Arg Met Ala Met Pro Arg Asp Phe Cys Ala Ile Ile His 65 70 75 80 Ala Pro Ala Val Ser Gly Pro Gly Pro His Val Met Leu Gly Leu Val 85 90 95 Asp Ser Gly Tyr Arg Gly Thr Val Leu Ala Val Val Val Ala Pro Asn 100 105 110 Gly Thr Arg Gly Phe Ala Pro Gly Ala Leu Arg Val Asp Val Thr Phe 115 120 125 Leu Asp Ile Arg Ala Thr Pro Pro Thr Leu Thr Glu Pro Ser Ser Leu 130 135 140 His Arg Phe Pro Gln Leu Ala Pro Ser Pro Leu Ala Gly Leu Arg Glu 145 150 155 160 Asp Pro Trp Leu Asp Gly Ala Leu Ala Thr Ala Gly Gly Ala Val Ala 165 170 175 Leu Pro Ala Arg Arg Arg Gly Gly Ser Leu Val Tyr Ala Gly Glu Leu 180 185 190 Thr Gln Val Thr Thr Glu His Gly Asp Cys Val His Glu Ala Pro Ala 195 200 205 Phe Leu Pro Lys Arg Glu Glu Asp Ala Gly Phe Asp Ile Leu Ile His 210 215 220 Arg Ala Val Thr Val Pro Ala Asn Gly Ala Thr Val Ile Gln Pro Ser 225 230 235 240 Leu Arg Val Leu Arg Ala Ala Asp Gly Pro Glu Ala Cys Tyr Val Leu 245 250 255 Gly Arg Ser Ser Leu Asn Ala Arg Gly Leu Leu Val Met Pro Thr Arg 260 265 270 Trp Pro Ser Gly His Ala Cys Ala Phe Val Val Cys Asn Leu Thr Gly 275 280 285 Val Pro Val Thr Leu Gln Ala Gly Ser Lys Val Ala Gln Leu Leu Val 290 295 300 Ala Gly Thr His Ala Leu Pro Trp Ile Pro Pro Asp Asn Ile His Glu 305 310 315 320 Asp Gly Ala Phe Arg Ala Tyr Pro Arg Gly Val Pro Asp Ala Thr Ala 325 330 335 Thr Pro Arg Asp Pro Pro Ile Leu Val Phe Thr Asn Glu Phe Asp Ala 340 345 350 Asp Ala Pro Pro Ser Lys Arg Gly Ala Gly Gly Phe Gly Ser Thr Gly 355 360 365 Ile <210> SEQ ID NO 154 <211> LENGTH: 532 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 154 Met Glu Leu Ser Tyr Ala Thr Thr Leu His His Arg Asp Val Val Phe 5 10 15 Tyr Val Thr Ala Asp Arg Asn Arg Ala Tyr Phe Val Cys Gly Gly Ser 20 25 30 Val Tyr Ser Val Gly Arg Pro Arg Asp Ser Gln Pro Gly Glu Ile Ala 35 40 45 Lys Phe Gly Leu Val Val Arg Gly Thr Gly Pro Lys Asp Arg Met Val 50 55 60 Ala Asn Tyr Val Arg Ser Glu Leu Arg Gln Arg Gly Leu Arg Asp Val 65 70 75 80 Arg Pro Val Gly Glu Asp Glu Val Phe Leu Asp Ser Val Cys Leu Leu 85 90 95 Asn Pro Asn Val Ser Ser Glu Arg Asp Val Ile Asn Thr Asn Asp Val 100 105 110 Glu Val Leu Asp Glu Cys Leu Ala Glu Tyr Cys Thr Ser Leu Arg Thr 115 120 125 Ser Pro Gly Val Leu Val Thr Gly Val Arg Val Arg Ala Arg Asp Arg 130 135 140 Val Ile Glu Leu Phe Glu His Pro Ala Ile Val Asn Ile Ser Ser Arg 145 150 155 160 Phe Ala Tyr Thr Pro Ser Pro Tyr Val Phe Ala Leu Ala Gln Ala His 165 170 175 Leu Pro Arg Leu Pro Ser Ser Leu Glu Pro Leu Val Ser Gly Leu Phe 180 185 190 Asp Gly Ile Pro Ala Pro Arg Gln Pro Leu Asp Ala Arg Asp Arg Arg 195 200 205 Thr Asp Val Val Ile Thr Gly Thr Arg Ala Pro Arg Pro Met Ala Gly 210 215 220 Thr Gly Ala Gly Gly Ala Gly Ala Lys Arg Ala Thr Val Ser Glu Phe 225 230 235 240 Val Gln Val Lys His Ile Asp Arg Val Val Ser Pro Ser Val Ser Ser 245 250 255 Ala Pro Pro Pro Ser Ala Pro Asp Ala Ser Leu Pro Pro Pro Gly Leu 260 265 270 Gln Glu Ala Ala Pro Pro Gly Pro Pro Leu Arg Glu Leu Trp Trp Val 275 280 285 Phe Tyr Ala Gly Asp Arg Ala Leu Glu Glu Pro His Ala Glu Ser Gly 290 295 300 Leu Thr Arg Glu Glu Val Arg Ala Val His Gly Phe Arg Glu Gln Ala 305 310 315 320 Trp Lys Leu Phe Gly Ser Val Gly Ala Pro Arg Ala Phe Leu Gly Ala 325 330 335 Ala Leu Ala Leu Ser Pro Thr Gln Lys Leu Ala Val Tyr Tyr Tyr Leu 340 345 350 Ile His Arg Glu Arg Arg Met Ser Pro Phe Pro Ala Leu Val Arg Leu 355 360 365 Val Gly Arg Tyr Ile Gln Arg His Gly Leu Tyr Val Pro Ala Pro Asp 370 375 380 Glu Pro Thr Leu Ala Asp Ala Met Asn Gly Leu Phe Arg Asp Ala Leu 385 390 395 400 Ala Ala Gly Thr Val Ala Glu Gln Leu Leu Met Phe Asp Leu Leu Pro 405 410 415 Pro Lys Asp Val Pro Val Gly Ser Asp Ala Arg Ala Asp Ser Ala Ala 420 425 430 Leu Leu Arg Phe Val Asp Ser Gln Arg Leu Thr Pro Gly Gly Ser Val 435 440 445 Ser Pro Glu His Val Met Tyr Leu Gly Ala Phe Leu Gly Val Leu Tyr 450 455 460 Ala Gly His Gly Arg Leu Ala Ala Ala Thr His Thr Ala Arg Leu Thr 465 470 475 480 Gly Val Thr Ser Leu Val Leu Thr Val Gly Asp Val Asp Arg Met Ser 485 490 495 Ala Phe Asp Arg Gly Pro Ala Gly Ala Ala Gly Arg Thr Arg Thr Ala 500 505 510 Gly Tyr Leu Asp Ala Leu Leu Thr Val Cys Leu Ala Arg Ala Gln His 515 520 525 Gly Gln Ser Val 530 <210> SEQ ID NO 155 <211> LENGTH: 481 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 155 Met Ala Cys Arg Lys Phe Cys Gly Val Tyr Arg Arg Pro Asp Lys Arg 5 10 15 Gln Glu Ala Ser Val Pro Pro Glu Thr Asn Thr Ala Pro Ala Phe Pro 20 25 30 Ala Ser Thr Phe Tyr Thr Pro Ala Glu Asp Ala Tyr Leu Ala Pro Gly 35 40 45 Pro Pro Glu Thr Ile His Pro Ser Arg Pro Pro Ser Pro Gly Glu Ala 50 55 60 Ala Arg Leu Cys Gln Leu Gln Glu Ile Leu Ala Gln Met His Ser Asp 65 70 75 80 Glu Asp Tyr Pro Ile Val Asp Ala Ala Gly Ala Glu Glu Glu Asp Glu 85 90 95 Ala Asp Asp Asp Ala Pro Asp Asp Val Ala Tyr Pro Glu Asp Tyr Ala 100 105 110 Glu Gly Arg Phe Leu Ser Met Val Ser Ala Ala Pro Leu Pro Gly Ala 115 120 125 Ser Gly His Pro Pro Val Pro Gly Arg Ala Ala Pro Pro Asp Val Arg 130 135 140 Thr Cys Asp Thr Gly Lys Val Gly Ala Thr Gly Phe Thr Pro Glu Glu 145 150 155 160 Leu Asp Thr Met Asp Arg Glu Ala Leu Arg Ala Ile Ser Arg Gly Cys 165 170 175 Lys Pro Pro Ser Thr Leu Ala Lys Leu Val Thr Gly Leu Gly Phe Ala 180 185 190 Ile His Gly Ala Leu Ile Pro Gly Ser Glu Gly Cys Val Phe Asp Ser 195 200 205 Ser His Pro Asn Tyr Pro His Arg Val Ile Val Lys Ala Gly Trp Tyr 210 215 220 Ala Ser Thr Ser His Glu Ala Arg Leu Leu Arg Arg Leu Asn His Pro 225 230 235 240 Ala Ile Leu Pro Leu Leu Asp Leu His Val Val Ser Gly Val Thr Cys 245 250 255 Leu Val Leu Pro Lys Tyr His Cys Asp Leu Tyr Thr Tyr Leu Ser Lys 260 265 270 Arg Pro Ser Pro Leu Gly His Leu Gln Ile Thr Ala Val Ser Arg Gln 275 280 285 Leu Leu Ser Ala Ile Asp Tyr Val His Cys Lys Gly Ile Ile His Arg 290 295 300 Asp Ile Lys Thr Glu Asn Ile Phe Ile Asn Thr Pro Glu Asn Ile Cys 305 310 315 320 Leu Gly Asp Phe Gly Ala Ala Cys Phe Val Arg Gly Cys Arg Ser Ser 325 330 335 Pro Phe His Tyr Gly Ile Ala Gly Thr Ile Asp Thr Asn Ala Pro Glu 340 345 350 Val Leu Ala Gly Asp Pro Tyr Thr Gln Val Ile Asp Ile Trp Ser Ala 355 360 365 Gly Leu Val Ile Phe Glu Thr Ala Val His Thr Ala Ser Leu Phe Ser 370 375 380 Ala Pro Arg Asp Pro Glu Arg Arg Pro Cys Asp Asn Gln Ile Ala Arg 385 390 395 400 Ile Ile Arg Gln Ala Gln Val His Val Asp Glu Phe Pro Thr His Ala 405 410 415 Glu Ser Arg Leu Thr Ala His Tyr Arg Ser Arg Ala Ala Gly Asn Asn 420 425 430 Arg Pro Ala Trp Thr Arg Pro Ala Trp Thr Arg Tyr Tyr Lys Ile His 435 440 445 Thr Asp Val Glu Tyr Leu Ile Cys Lys Ala Leu Thr Phe Asp Ala Ala 450 455 460 Leu Arg Pro Ser Ala Ala Glu Leu Leu Arg Leu Pro Leu Phe His Pro 465 470 475 480 Lys <210> SEQ ID NO 156 <211> LENGTH: 512 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 156 Met Ala Thr Asp Ile Asp Met Leu Ile Asp Leu Gly Leu Asp Leu Ser 5 10 15 Asp Ser Glu Leu Glu Glu Asp Ala Leu Glu Arg Asp Glu Glu Gly Arg 20 25 30 Arg Asp Asp Pro Glu Ser Asp Ser Ser Gly Glu Cys Ser Ser Ser Asp 35 40 45 Glu Asp Met Glu Asp Pro Cys Gly Asp Gly Gly Ala Glu Ala Ile Asp 50 55 60 Ala Ala Ile Pro Lys Gly Pro Pro Ala Arg Pro Glu Asp Ala Gly Thr 65 70 75 80 Pro Glu Ala Ser Thr Pro Arg Pro Ala Ala Arg Arg Gly Ala Asp Asp 85 90 95 Pro Pro Pro Ala Thr Thr Gly Val Trp Ser Arg Leu Gly Thr Arg Arg 100 105 110 Ser Ala Ser Pro Arg Glu Pro His Gly Gly Lys Val Ala Arg Ile Gln 115 120 125 Pro Pro Ser Thr Lys Ala Pro His Pro Arg Gly Gly Arg Arg Gly Arg 130 135 140 Arg Arg Gly Arg Gly Arg Tyr Gly Pro Gly Gly Ala Asp Ser Thr Pro 145 150 155 160 Lys Pro Arg Arg Arg Val Ser Arg Asn Ala His Asn Gln Gly Gly Arg 165 170 175 His Pro Ala Ser Ala Arg Thr Asp Gly Pro Gly Ala Thr His Gly Glu 180 185 190 Ala Arg Arg Gly Gly Glu Gln Leu Asp Val Ser Gly Gly Pro Arg Pro 195 200 205 Arg Gly Thr Arg Gln Ala Pro Pro Pro Leu Met Ala Leu Ser Leu Thr 210 215 220 Pro Pro His Ala Asp Gly Arg Ala Pro Val Pro Glu Arg Lys Ala Pro 225 230 235 240 Ser Ala Asp Thr Ile Asp Pro Ala Val Arg Ala Val Leu Arg Ser Ile 245 250 255 Ser Glu Arg Ala Ala Val Glu Arg Ile Ser Glu Ser Phe Gly Arg Ser 260 265 270 Ala Leu Val Met Gln Asp Pro Phe Gly Gly Met Pro Phe Pro Ala Ala 275 280 285 Asn Ser Pro Trp Ala Pro Val Leu Ala Thr Gln Ala Gly Gly Phe Asp 290 295 300 Ala Glu Thr Arg Arg Val Ser Trp Glu Thr Leu Val Ala His Gly Pro 305 310 315 320 Ser Leu Tyr Arg Thr Phe Ala Ala Asn Pro Arg Ala Ala Ser Thr Ala 325 330 335 Lys Ala Met Arg Asp Cys Val Leu Arg Gln Glu Asn Leu Ile Glu Ala 340 345 350 Leu Ala Ser Ala Asp Glu Thr Leu Ala Trp Cys Lys Met Cys Ile His 355 360 365 His Asn Leu Pro Leu Arg Pro Gln Asp Pro Ile Ile Gly Thr Ala Ala 370 375 380 Ala Val Leu Glu Asn Leu Ala Thr Arg Leu Arg Pro Phe Leu Gln Cys 385 390 395 400 Tyr Leu Lys Ala Arg Gly Leu Cys Gly Leu Asp Asp Leu Cys Ser Arg 405 410 415 Arg Arg Leu Ser Asp Ile Lys Asp Ile Ala Ser Phe Val Leu Val Ile 420 425 430 Leu Ala Arg Leu Ala Asn Arg Val Glu Arg Gly Val Ser Glu Ile Asp 435 440 445 Tyr Thr Thr Val Gly Val Gly Ala Gly Glu Thr Met His Phe Tyr Ile 450 455 460 Pro Gly Ala Cys Met Ala Gly Leu Ile Glu Ile Leu Asp Thr His Arg 465 470 475 480 Gln Glu Cys Ser Ser Arg Val Cys Glu Leu Thr Ala Ser His Thr Ile 485 490 495 Ala Pro Leu Tyr Val His Gly Lys Tyr Phe Tyr Cys Asn Ser Leu Phe 500 505 510 <210> SEQ ID NO 157 <211> LENGTH: 545 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 157 Met Ala Arg Gly Ala Gly Leu Val Phe Phe Val Gly Val Trp Val Val 5 10 15 Ser Cys Leu Ala Ala Ala Pro Arg Thr Ser Trp Lys Arg Val Thr Ser 20 25 30 Gly Glu Asp Val Val Leu Leu Pro Ala Pro Ala Glu Arg Thr Arg Ala 35 40 45 His Lys Leu Leu Trp Ala Ala Glu Pro Leu Asp Ala Cys Gly Pro Leu 50 55 60 Arg Pro Ser Trp Val Ala Leu Trp Pro Pro Arg Arg Val Leu Glu Thr 65 70 75 80 Val Val Asp Ala Ala Cys Met Arg Ala Pro Glu Pro Leu Ala Ile Ala 85 90 95 Tyr Ser Pro Pro Phe Pro Ala Gly Asp Glu Gly Leu Tyr Ser Glu Leu 100 105 110 Ala Trp Arg Asp Arg Val Ala Val Val Asn Glu Ser Leu Val Ile Tyr 115 120 125 Gly Ala Leu Glu Thr Asp Ser Gly Leu Tyr Thr Leu Ser Val Val Gly 130 135 140 Leu Ser Asp Glu Ala Arg Gln Val Ala Ser Val Val Leu Val Val Glu 145 150 155 160 Pro Ala Pro Val Pro Thr Pro Thr Pro Asp Asp Tyr Asp Glu Glu Asp 165 170 175 Asp Ala Gly Val Thr Asn Ala Arg Arg Ser Ala Phe Pro Pro Gln Pro 180 185 190 Pro Pro Arg Arg Pro Pro Val Ala Pro Pro Thr His Pro Arg Val Ile 195 200 205 Pro Glu Val Ser His Val Arg Gly Val Thr Val His Met Glu Thr Leu 210 215 220 Glu Ala Ile Leu Phe Ala Pro Gly Glu Thr Phe Gly Thr Asn Val Ser 225 230 235 240 Ile His Ala Ile Ala His Asp Asp Gly Pro Tyr Ala Met Asp Val Val 245 250 255 Trp Met Arg Phe Asp Val Pro Ser Ser Cys Ala Asp Met Arg Ile Tyr 260 265 270 Glu Ala Cys Leu Tyr His Pro Gln Leu Pro Glu Cys Leu Ser Pro Ala 275 280 285 Asp Ala Pro Cys Ala Val Ser Ser Trp Ala Tyr Arg Leu Ala Val Arg 290 295 300 Ser Tyr Ala Gly Cys Ser Arg Thr Thr Pro Pro Pro Arg Cys Phe Ala 305 310 315 320 Glu Ala Arg Met Glu Pro Val Pro Gly Leu Ala Trp Leu Ala Ser Thr 325 330 335 Val Asn Leu Glu Phe Gln His Ala Ser Pro Gln His Ala Gly Leu Tyr 340 345 350 Leu Cys Val Val Tyr Val Asp Asp His Ile His Ala Trp Gly His Met 355 360 365 Thr Ile Ser Thr Ala Ala Gln Tyr Arg Asn Ala Val Val Glu Gln His 370 375 380 Leu Pro Gln Arg Gln Pro Glu Pro Val Glu Pro Thr Arg Pro His Val 385 390 395 400 Arg Ala Pro His Pro Ala Pro Ser Ala Arg Gly Pro Leu Arg Leu Gly 405 410 415 Ala Val Leu Gly Ala Ala Leu Leu Leu Ala Ala Leu Gly Leu Ser Ala 420 425 430 Trp Ala Cys Met Thr Cys Trp Arg Arg Arg Ser Trp Arg Ala Val Lys 435 440 445 Ser Arg Ala Ser Ala Thr Gly Pro Thr Tyr Ile Arg Val Ala Asp Ser 450 455 460 Glu Leu Tyr Ala Asp Trp Ser Ser Asp Ser Glu Gly Glu Arg Asp Gly 465 470 475 480 Ser Leu Trp Gln Asp Pro Pro Glu Arg Pro Asp Ser Pro Ser Thr Asn 485 490 495 Gly Ser Gly Phe Glu Ile Leu Ser Pro Thr Ala Pro Ser Val Tyr Pro 500 505 510 His Ser Glu Gly Arg Lys Ser Arg Arg Pro Leu Thr Thr Phe Gly Ser 515 520 525 Gly Ser Pro Gly Arg Arg His Ser Gln Ala Ser Tyr Pro Ser Val Leu 530 535 540 Trp 545 <210> SEQ ID NO 158 <211> LENGTH: 1374 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 158 Met Ala Ala Pro Ala Arg Asp Pro Pro Gly Tyr Arg Tyr Ala Ala Ala 5 10 15 Ile Leu Pro Thr Gly Ser Ile Leu Ser Thr Ile Glu Val Ala Ser His 20 25 30 Arg Arg Leu Phe Asp Phe Phe Ala Ala Val Arg Ser Asp Glu Asn Ser 35 40 45 Leu Tyr Asp Val Glu Phe Asp Ala Leu Leu Gly Ser Tyr Cys Asn Thr 50 55 60 Leu Ser Leu Val Arg Phe Leu Glu Leu Gly Leu Ser Val Ala Cys Val 65 70 75 80 Cys Thr Lys Phe Pro Glu Leu Ala Tyr Met Asn Glu Gly Arg Val Gln 85 90 95 Phe Glu Val His Gln Pro Leu Ile Ala Arg Asp Gly Pro His Pro Val 100 105 110 Glu Gln Pro Val His Asn Tyr Met Thr Lys Val Ile Asp Arg Arg Ala 115 120 125 Leu Asn Ala Ala Phe Ser Leu Ala Thr Glu Ala Ile Ala Leu Leu Thr 130 135 140 Gly Glu Ala Leu Asp Gly Thr Gly Ile Ser Leu His Arg Gln Leu Arg 145 150 155 160 Ala Ile Gln Gln Leu Ala Arg Asn Val Gln Ala Val Leu Gly Ala Phe 165 170 175 Glu Arg Gly Thr Ala Asp Gln Met Leu His Val Leu Leu Glu Lys Ala 180 185 190 Pro Pro Leu Ala Leu Leu Leu Pro Met Gln Arg Tyr Leu Asp Asn Gly 195 200 205 Arg Leu Ala Thr Arg Val Ala Arg Ala Thr Leu Val Ala Glu Leu Lys 210 215 220 Arg Ser Phe Cys Asp Thr Ser Phe Phe Leu Gly Lys Ala Gly His Arg 225 230 235 240 Arg Glu Ala Ile Glu Ala Trp Leu Val Asp Leu Thr Thr Ala Thr Gln 245 250 255 Pro Ser Val Ala Val Pro Arg Leu Thr His Ala Asp Thr Arg Gly Arg 260 265 270 Pro Val Asp Gly Val Leu Val Thr Thr Ala Ala Ile Lys Gln Arg Leu 275 280 285 Leu Gln Ser Phe Leu Lys Val Glu Asp Thr Glu Ala Asp Val Pro Val 290 295 300 Thr Tyr Gly Glu Met Val Leu Asn Gly Ala Asn Leu Val Thr Ala Leu 305 310 315 320 Val Met Gly Lys Ala Val Arg Ser Leu Asp Asp Val Gly Arg His Leu 325 330 335 Leu Asp Met Gln Glu Glu Gln Leu Glu Ala Asn Arg Glu Thr Leu Asp 340 345 350 Glu Leu Glu Ser Ala Pro Gln Thr Thr Arg Val Arg Ala Asp Leu Val 355 360 365 Ala Ile Gly Asp Arg Leu Val Phe Leu Glu Ala Leu Glu Arg Arg Ile 370 375 380 Tyr Ala Ala Thr Asn Val Pro Tyr Pro Leu Val Gly Ala Met Asp Leu 385 390 395 400 Thr Phe Val Leu Pro Leu Gly Leu Phe Asn Pro Ala Met Glu Arg Phe 405 410 415 Ala Ala His Ala Gly Asp Leu Val Pro Ala Pro Gly His Pro Glu Pro 420 425 430 Arg Ala Phe Pro Pro Arg Gln Leu Phe Phe Trp Gly Lys Asp His Gln 435 440 445 Val Leu Arg Leu Ser Met Glu Asn Ala Val Gly Thr Val Cys His Pro 450 455 460 Ser Leu Met Asn Ile Asp Ala Ala Val Gly Gly Val Asn His Asp Pro 465 470 475 480 Val Glu Ala Ala Asn Pro Tyr Gly Ala Tyr Val Ala Ala Pro Ala Gly 485 490 495 Pro Gly Ala Asp Met Gln Gln Arg Phe Leu Asn Ala Trp Arg Gln Arg 500 505 510 Leu Ala His Gly Arg Val Arg Trp Val Ala Glu Cys Gln Met Thr Ala 515 520 525 Glu Gln Phe Met Gln Pro Asp Asn Ala Asn Leu Ala Leu Glu Leu His 530 535 540 Pro Ala Phe Asp Phe Phe Ala Gly Val Ala Asp Val Glu Leu Pro Gly 545 550 555 560 Gly Glu Val Pro Pro Ala Gly Pro Gly Ala Ile Gln Ala Thr Trp Arg 565 570 575 Val Val Asn Gly Asn Leu Pro Leu Ala Leu Cys Pro Val Ala Phe Arg 580 585 590 Asp Ala Arg Gly Leu Glu Leu Gly Val Gly Arg His Ala Met Ala Pro 595 600 605 Ala Thr Ile Ala Ala Val Arg Gly Ala Phe Glu Asp Arg Ser Tyr Pro 610 615 620 Ala Val Phe Tyr Leu Leu Gln Ala Ala Ile His Gly Asn Glu His Val 625 630 635 640 Phe Cys Ala Leu Ala Arg Leu Val Thr Gln Cys Ile Thr Ser Tyr Trp 645 650 655 Asn Asn Thr Arg Cys Ala Ala Phe Val Asn Asp Tyr Ser Leu Val Ser 660 665 670 Tyr Ile Val Thr Tyr Leu Gly Gly Asp Leu Pro Glu Glu Cys Met Ala 675 680 685 Val Tyr Arg Asp Leu Val Ala His Val Glu Ala Leu Ala Gln Leu Val 690 695 700 Asp Asp Phe Thr Leu Pro Gly Pro Glu Leu Gly Gly Gln Ala Gln Ala 705 710 715 720 Glu Leu Asn His Leu Met Arg Asp Pro Ala Leu Leu Pro Pro Leu Val 725 730 735 Trp Asp Cys Asp Gly Leu Met Arg His Ala Ala Leu Asp Arg His Arg 740 745 750 Asp Cys Arg Ile Asp Ala Gly Gly His Glu Pro Val Tyr Ala Ala Ala 755 760 765 Cys Asn Val Ala Thr Ala Asp Phe Asn Arg Asn Asp Gly Arg Leu Leu 770 775 780 His Asn Thr Gln Ala Arg Ala Ala Asp Ala Ala Asp Asp Arg Pro His 785 790 795 800 Arg Pro Ala Asp Trp Thr Val His His Lys Ile Tyr Tyr Tyr Val Leu 805 810 815 Val Pro Ala Phe Ser Arg Gly Arg Cys Cys Thr Ala Gly Val Arg Phe 820 825 830 Asp Arg Val Tyr Ala Thr Leu Gln Asn Met Val Val Pro Glu Ile Ala 835 840 845 Pro Gly Glu Glu Cys Pro Ser Asp Pro Val Thr Asp Pro Ala His Pro 850 855 860 Leu His Pro Ala Asn Leu Val Ala Asn Thr Val Lys Arg Met Phe His 865 870 875 880 Asn Gly Arg Val Val Val Asp Gly Pro Ala Met Leu Thr Leu Gln Val 885 890 895 Leu Ala His Asn Met Ala Glu Arg Thr Thr Ala Leu Leu Cys Ser Ala 900 905 910 Ala Pro Asp Ala Gly Ala Asn Thr Ala Ser Thr Ala Asn Met Arg Ile 915 920 925 Phe Asp Gly Ala Leu His Ala Gly Val Leu Leu Met Ala Pro Gln His 930 935 940 Leu Asp His Thr Ile Gln Asn Gly Glu Tyr Phe Tyr Val Leu Pro Val 945 950 955 960 His Ala Leu Phe Ala Gly Ala Asp His Val Ala Asn Ala Pro Asn Phe 965 970 975 Pro Pro Ala Leu Arg Asp Leu Ala Arg Asp Val Pro Leu Val Pro Pro 980 985 990 Ala Leu Gly Ala Asn Tyr Phe Ser Ser Ile Arg Gln Pro Val Val Gln 995 1000 1005 His Ala Arg Glu Ser Ala Ala Gly Glu Asn Ala Leu Thr Tyr Ala Leu 1010 1015 1020 Met Ala Gly Tyr Phe Lys Met Ser Pro Val Ala Leu Tyr His Gln Leu 1025 1030 1035 1040 Lys Thr Gly Leu His Pro Gly Phe Gly Phe Thr Val Val Arg Gln Asp 1045 1050 1055 Arg Phe Val Thr Glu Asn Val Leu Phe Ser Glu Arg Ala Ser Glu Ala 1060 1065 1070 Tyr Phe Leu Gly Gln Leu Gln Val Ala Arg His Glu Thr Gly Gly Gly 1075 1080 1085 Val Asn Phe Thr Leu Thr Gln Pro Arg Gly Asn Val Asp Leu Gly Val 1090 1095 1100 Gly Tyr Thr Ala Val Ala Ala Thr Gly Thr Val Arg Asn Pro Val Thr 1105 1110 1115 1120 Asp Met Gly Asn Leu Pro Gln Asn Phe Tyr Leu Gly Arg Gly Ala Pro 1125 1130 1135 Pro Leu Leu Asp Asn Ala Ala Ala Val Tyr Leu Arg Asn Ala Val Val 1140 1145 1150 Ala Gly Asn Arg Leu Gly Pro Ala Gln Pro Leu Pro Val Phe Gly Cys 1155 1160 1165 Ala Gln Val Pro Arg Arg Ala Gly Met Asp His Gly Gln Asp Ala Val 1170 1175 1180 Cys Glu Phe Ile Ala Thr Pro Val Ala Thr Asp Ile Asn Tyr Phe Arg 1185 1190 1195 1200 Arg Pro Cys Asn Pro Arg Gly Arg Ala Ala Gly Gly Val Tyr Ala Gly 1205 1210 1215 Asp Lys Glu Gly Asp Val Ile Ala Leu Met Tyr Asp His Gly Gln Ser 1220 1225 1230 Asp Pro Ala Arg Pro Phe Ala Ala Thr Ala Asn Pro Trp Ala Ser Gln 1235 1240 1245 Arg Phe Ser Tyr Gly Asp Leu Leu Tyr Asn Gly Ala Tyr His Leu Asn 1250 1255 1260 Gly Ala Ser Pro Val Leu Ser Pro Cys Phe Lys Phe Phe Thr Ala Ala 1265 1270 1275 1280 Asp Ile Thr Ala Lys His Arg Cys Leu Glu Arg Leu Ile Val Glu Thr 1285 1290 1295 Gly Ser Ala Val Ser Thr Ala Thr Ala Ala Ser Asp Val Gln Phe Lys 1300 1305 1310 Arg Pro Pro Gly Cys Arg Glu Leu Val Glu Asp Pro Cys Gly Leu Phe 1315 1320 1325 Gln Glu Ala Tyr Pro Ile Thr Cys Ala Ser Asp Pro Ala Leu Leu Arg 1330 1335 1340 Ser Ala Arg Asp Gly Glu Ala His Ala Arg Glu Thr His Phe Thr Gln 1345 1350 1355 1360 Tyr Leu Ile Tyr Asp Ala Ser Pro Leu Lys Gly Leu Ser Leu 1365 1370 <210> SEQ ID NO 159 <211> LENGTH: 722 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 159 Met Gln Arg Arg Ala Arg Gly Ala Ser Ser Leu Arg Leu Ala Arg Cys 5 10 15 Leu Thr Pro Ala Asn Leu Ile Arg Gly Ala Asn Ala Gly Val Pro Glu 20 25 30 Arg Arg Ile Phe Ala Gly Cys Leu Leu Pro Thr Pro Glu Gly Leu Leu 35 40 45 Ser Ala Ala Val Gly Val Leu Arg Gln Arg Ala Asp Asp Leu Gln Pro 50 55 60 Ala Phe Leu Thr Gly Ala Asp Arg Ser Val Arg Leu Ala Ala Arg His 65 70 75 80 His Asn Thr Val Pro Glu Ser Leu Ile Val Asp Gly Leu Ala Ser Asp 85 90 95 Pro His Tyr Asp Tyr Ile Arg His Tyr Ala Ser Ala Ala Lys Gln Ala 100 105 110 Leu Gly Glu Val Glu Leu Ser Gly Gly Gln Leu Ser Arg Ala Ile Leu 115 120 125 Ala Gln Tyr Trp Lys Tyr Leu Gln Thr Val Val Pro Ser Gly Leu Asp 130 135 140 Ile Pro Asp Asp Pro Ala Gly Asp Cys Asp Pro Ser Leu His Val Leu 145 150 155 160 Leu Arg Pro Thr Leu Leu Pro Lys Leu Leu Val Arg Ala Pro Phe Lys 165 170 175 Ser Gly Ala Ala Ala Ala Lys Tyr Ala Ala Ala Val Ala Gly Leu Arg 180 185 190 Asp Ala Ala His Arg Leu Gln Gln Tyr Met Phe Phe Met Arg Pro Ala 195 200 205 Asp Pro Ser Arg Pro Ser Thr Asp Thr Ala Leu Arg Leu Ser Glu Leu 210 215 220 Leu Ala Tyr Val Ser Val Leu Tyr His Trp Ala Ser Trp Met Leu Trp 225 230 235 240 Thr Ala Asp Lys Tyr Val Cys Arg Arg Leu Gly Pro Ala Asp Arg Arg 245 250 255 Phe Val Ala Leu Ser Gly Ser Leu Glu Ala Pro Ala Glu Thr Phe Ala 260 265 270 Arg His Leu Asp Arg Gly Pro Ser Gly Thr Thr Gly Ser Met Gln Cys 275 280 285 Met Ala Leu Arg Ala Ala Val Ser Asp Val Leu Gly His Leu Thr Arg 290 295 300 Leu Ala His Leu Trp Glu Thr Gly Lys Arg Ser Gly Gly Thr Tyr Gly 305 310 315 320 Ile Val Asp Ala Ile Val Ser Thr Val Glu Val Leu Ser Ile Val His 325 330 335 His His Ala Gln Tyr Ile Ile Asn Ala Thr Leu Thr Gly Tyr Val Val 340 345 350 Trp Ala Ser Asp Ser Leu Asn Asn Glu Tyr Leu Thr Ala Ala Val Asp 355 360 365 Ser Gln Glu Arg Phe Cys Arg Thr Ala Ala Pro Leu Phe Pro Thr Met 370 375 380 Thr Ala Pro Ser Trp Ala Arg Met Glu Leu Ser Ile Lys Ser Trp Phe 385 390 395 400 Gly Ala Ala Leu Ala Pro Asp Leu Leu Arg Ser Gly Thr Pro Ser Pro 405 410 415 His Tyr Glu Ser Ile Leu Arg Leu Ala Ala Ser Gly Pro Pro Gly Gly 420 425 430 Arg Gly Ala Val Gly Gly Ser Cys Arg Asp Lys Ile Gln Arg Thr Arg 435 440 445 Arg Asp Asn Ala Pro Pro Pro Leu Pro Arg Ala Arg Pro His Ser Thr 450 455 460 Pro Ala Ala Pro Arg Arg Cys Arg Arg His Arg Glu Asp Leu Pro Glu 465 470 475 480 Pro Pro His Val Asp Ala Ala Asp Arg Gly Pro Glu Pro Cys Ala Gly 485 490 495 Arg Pro Ala Thr Tyr Tyr Thr His Met Ala Gly Ala Pro Pro Arg Leu 500 505 510 Pro Pro Arg Asn Pro Ala Pro Pro Glu Gln Arg Pro Ala Ala Ala Ala 515 520 525 Arg Pro Leu Ala Ala Gln Arg Glu Ala Ala Gly Val Tyr Asp Ala Val 530 535 540 Arg Thr Trp Gly Pro Asp Ala Glu Ala Glu Pro Asp Gln Met Glu Asn 545 550 555 560 Thr Tyr Leu Leu Pro Asp Asp Asp Ala Ala Met Pro Ala Gly Val Gly 565 570 575 Leu Gly Ala Thr Pro Ala Ala Asp Thr Thr Ala Ala Ala Ala Trp Pro 580 585 590 Ala Glu Ser His Ala Pro Arg Ala Pro Ser Glu Asp Ala Asp Ser Ile 595 600 605 Tyr Glu Ser Val Gly Glu Asp Gly Gly Arg Val Tyr Glu Glu Ile Pro 610 615 620 Trp Val Arg Val Tyr Glu Asn Ile Cys Pro Arg Arg Arg Leu Ala Gly 625 630 635 640 Gly Ala Ala Leu Pro Gly Asp Ala Pro Asp Ser Pro Tyr Ile Glu Ala 645 650 655 Glu Asn Pro Leu Tyr Asp Trp Gly Gly Ser Ala Leu Phe Ser Pro Arg 660 665 670 Arg Ala Thr Arg Ala Pro Asp Pro Gly Leu Ser Leu Ser Pro Met Pro 675 680 685 Ala Arg Pro Arg Thr Asn Ala Leu Ala Asn Asp Gly Pro Thr Asn Val 690 695 700 Ala Ala Leu Ser Ala Leu Leu Thr Lys Leu Lys Arg Gly Arg His Gln 705 710 715 720 Ser His <210> SEQ ID NO 160 <211> LENGTH: 318 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 160 Met Ile Thr Asp Cys Phe Glu Ala Asp Ile Ala Ile Pro Ser Gly Ile 5 10 15 Ser Arg Pro Asp Ala Ala Ala Leu Gln Arg Cys Glu Gly Arg Val Val 20 25 30 Phe Leu Pro Thr Ile Arg Arg Gln Leu Ala Leu Ala Asp Val Ala His 35 40 45 Glu Ser Phe Val Ser Gly Gly Val Ser Pro Asp Thr Leu Gly Leu Leu 50 55 60 Leu Ala Tyr Arg Arg Arg Phe Pro Ala Val Ile Thr Arg Val Leu Pro 65 70 75 80 Thr Arg Ile Val Ala Cys Pro Val Asp Leu Gly Leu Thr His Ala Gly 85 90 95 Thr Val Asn Leu Arg Asn Thr Ser Pro Val Asp Leu Cys Asn Gly Asp 100 105 110 Pro Val Ser Leu Val Pro Pro Val Phe Glu Gly Gln Ala Thr Asp Val 115 120 125 Arg Leu Glu Ser Leu Asp Leu Thr Leu Arg Phe Pro Val Pro Leu Pro 130 135 140 Thr Pro Leu Ala Arg Glu Ile Val Ala Arg Leu Val Ala Arg Gly Ile 145 150 155 160 Arg Asp Leu Asn Pro Asp Pro Arg Thr Pro Gly Glu Leu Pro Asp Leu 165 170 175 Asn Val Leu Tyr Tyr Asn Gly Ala Arg Leu Ser Leu Val Ala Asp Val 180 185 190 Gln Gln Leu Ala Ser Val Asn Thr Glu Leu Arg Ser Leu Val Leu Asn 195 200 205 Met Val Tyr Ser Ile Thr Glu Gly Thr Thr Leu Ile Leu Thr Leu Ile 210 215 220 Pro Arg Leu Leu Ala Leu Ser Ala Gln Asp Gly Tyr Val Asn Ala Leu 225 230 235 240 Leu Gln Met Gln Ser Val Thr Arg Glu Ala Ala Gln Leu Ile His Pro 245 250 255 Glu Ala Pro Met Leu Met Gln Asp Gly Glu Arg Arg Leu Pro Leu Tyr 260 265 270 Glu Ala Leu Val Ala Trp Leu Ala His Ala Gly Gln Leu Gly Asp Ile 275 280 285 Leu Ala Leu Ala Pro Ala Val Arg Val Cys Thr Phe Asp Gly Ala Ala 290 295 300 Val Val Gln Ser Gly Asp Met Ala Pro Val Ile Arg Tyr Pro 305 310 315 <210> SEQ ID NO 161 <211> LENGTH: 825 <212> TYPE: PRT <213> ORGANISM: HSV2 <400> SEQUENCE: 161 Met Glu Pro Arg Pro Gly Thr Ser Ser Arg Ala Asp Pro Gly Pro Glu 5 10 15 Arg Pro Pro Arg Gln Thr Pro Gly Thr Gln Pro Ala Ala Pro His Ala 20 25 30 Trp Gly Met Leu Asn Asp Met Gln Trp Leu Ala Ser Ser Asp Ser Glu 35 40 45 Glu Glu Thr Glu Val Gly Ile Ser Asp Asp Asp Leu His Arg Asp Ser 50 55 60 Thr Ser Glu Ala Gly Ser Thr Asp Thr Glu Met Phe Glu Ala Gly Leu 65 70 75 80 Met Asp Ala Ala Thr Pro Pro Ala Arg Pro Pro Ala Glu Arg Gln Gly 85 90 95 Ser Pro Thr Pro Ala Asp Ala Gln Gly Ser Cys Gly Gly Gly Pro Val 100 105 110 Gly Glu Glu Glu Ala Glu Ala Gly Gly Gly Gly Asp Val Cys Ala Val 115 120 125 Cys Thr Asp Glu Ile Ala Pro Pro Leu Arg Cys Gln Ser Phe Pro Cys 130 135 140 Leu His Pro Phe Cys Ile Pro Cys Met Lys Thr Trp Ile Pro Leu Arg 145 150 155 160 Asn Thr Cys Pro Leu Cys Asn Thr Pro Val Ala Tyr Leu Ile Val Gly 165 170 175 Val Thr Ala Ser Gly Ser Phe Ser Thr Ile Pro Ile Val Asn Asp Pro 180 185 190 Arg Thr Arg Val Glu Ala Glu Ala Ala Val Arg Ala Gly Thr Ala Val 195 200 205 Asp Phe Ile Trp Thr Gly Asn Pro Arg Thr Ala Pro Arg Ser Leu Ser 210 215 220 Leu Gly Gly His Thr Val Arg Ala Leu Ser Pro Thr Pro Pro Trp Pro 225 230 235 240 Gly Thr Asp Asp Glu Asp Asp Asp Leu Ala Asp Val Asp Tyr Val Pro 245 250 255 Pro Ala Pro Arg Arg Ala Pro Arg Arg Gly Gly Gly Gly Ala Gly Ala 260 265 270 Thr Arg Gly Thr Ser Gln Pro Ala Ala Thr Arg Pro Ala Pro Pro Gly 275 280 285 Ala Pro Arg Ser Ser Ser Ser Gly Gly Ala Pro Leu Arg Ala Gly Val 290 295 300 Gly Ser Gly Ser Gly Gly Gly Pro Ala Val Ala Ala Val Val Pro Arg 305 310 315 320 Val Ala Ser Leu Pro Pro Ala Ala Gly Gly Gly Arg Ala Gln Ala Arg 325 330 335 Arg Val Gly Glu Asp Ala Ala Ala Ala Glu Gly Arg Thr Pro Pro Ala 340 345 350 Arg Gln Pro Arg Ala Ala Gln Glu Pro Pro Ile Val Ile Ser Asp Ser 355 360 365 Pro Pro Pro Ser Pro Arg Arg Pro Ala Gly Pro Gly Pro Leu Ser Phe 370 375 380 Val Ser Ser Ser Ser Ala Gln Val Ser Ser Gly Pro Gly Gly Gly Gly 385 390 395 400 Leu Pro Gln Ser Ser Gly Arg Ala Ala Arg Pro Arg Ala Ala Val Ala 405 410 415 Pro Arg Val Arg Ser Pro Pro Arg Ala Ala Ala Ala Pro Val Val Ser 420 425 430 Ala Ser Ala Asp Ala Ala Gly Pro Ala Pro Pro Ala Val Pro Val Asp 435 440 445 Ala His Arg Ala Pro Arg Ser Arg Met Thr Gln Ala Gln Thr Asp Thr 450 455 460 Gln Ala Gln Ser Leu Gly Arg Ala Gly Ala Thr Asp Ala Arg Gly Ser 465 470 475 480 Gly Gly Pro Gly Ala Glu Gly Gly Pro Gly Val Pro Arg Gly Thr Asn 485 490 495 Thr Pro Gly Ala Ala Pro His Ala Ala Glu Gly Ala Ala Ala Arg Pro 500 505 510 Arg Lys Arg Arg Gly Ser Asp Ser Gly Pro Ala Ala Ser Ser Ser Ala 515 520 525 Ser Ser Ser Ala Ala Pro Arg Ser Pro Leu Ala Pro Gln Gly Val Gly 530 535 540 Ala Lys Arg Ala Ala Pro Arg Arg Ala Pro Asp Ser Asp Ser Gly Asp 545 550 555 560 Arg Gly His Gly Pro Leu Ala Pro Ala Ser Ala Gly Ala Ala Pro Pro 565 570 575 Ser Ala Ser Pro Ser Ser Gln Ala Ala Val Ala Ala Ala Ser Ser Ser 580 585 590 Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser 595 600 605 Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala Ser Ser Ser Ser Ala 610 615 620 Ser Ser Ser Ala Gly Gly Ala Gly Gly Ser Val Ala Ser Ala Ser Gly 625 630 635 640 Ala Gly Glu Arg Arg Glu Thr Ser Leu Gly Pro Arg Ala Ala Ala Pro 645 650 655 Arg Gly Pro Arg Lys Cys Ala Arg Lys Thr Arg His Ala Glu Gly Gly 660 665 670 Pro Glu Pro Gly Ala Arg Asp Pro Ala Pro Gly Leu Thr Arg Tyr Leu 675 680 685 Pro Ile Ala Gly Val Ser Ser Val Val Ala Leu Ala Pro Tyr Val Asn 690 695 700 Lys Thr Val Thr Gly Asp Cys Leu Pro Val Leu Asp Met Glu Thr Gly 705 710 715 720 His Ile Gly Ala Tyr Val Val Leu Val Asp Gln Thr Gly Asn Val Ala 725 730 735 Asp Leu Leu Arg Ala Ala Ala Pro Ala Trp Ser Arg Arg Thr Leu Leu 740 745 750 Pro Glu His Ala Arg Asn Cys Val Arg Pro Pro Asp Tyr Pro Thr Pro 755 760 765 Pro Ala Ser Glu Trp Asn Ser Leu Trp Met Thr Pro Val Gly Asn Met 770 775 780 Leu Phe Asp Gln Gly Thr Leu Val Gly Ala Leu Asp Phe His Gly Leu 785 790 795 800 Arg Ser Arg His Pro Trp Ser Arg Glu Gln Gly Ala Pro Ala Pro Ala 805 810 815 Gly Asp Ala Pro Ala Gly His Gly Glu 820 825 <210> SEQ ID NO 162 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 162 Ile Trp Thr Gly Asn Pro Arg Thr Ala Pro Arg Ser Leu Ser Leu 1 5 10 15 <210> SEQ ID NO 163 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 163 Tyr Met Phe Phe Met Arg Pro Ala Asp Pro Ser Arg Pro Ser Thr 1 5 10 15 <210> SEQ ID NO 164 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 164 Val Cys Arg Arg Leu Gly Pro Ala Asp Arg Arg Phe Val Ala Leu 1 5 10 15 <210> SEQ ID NO 165 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 165 Gly Pro Ala Asp Arg Arg Phe Val Ala Leu Ser Gly Ser Leu Glu 1 5 10 15 <210> SEQ ID NO 166 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 166 Ser Asp Val Leu Gly His Leu Thr Arg Leu Ala His Leu Trp Glu 1 5 10 15 <210> SEQ ID NO 167 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 167 Gly His Met Thr Ile Ser Thr Ala Ala Gln Tyr Arg Asn Ala Val 1 5 10 15 <210> SEQ ID NO 168 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 168 Leu Asn Ala Trp Arg Gln Arg Leu Ala His Gly Arg Val Arg Trp 1 5 10 15 <210> SEQ ID NO 169 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 169 Gln Arg Leu Ala His Gly Arg Val Arg Trp Val Ala Glu Cys Gln 1 5 10 15 <210> SEQ ID NO 170 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 170 Asp Leu Val Ala Ile Gly Asp Arg Leu Val Phe Leu Glu Ala Leu 1 5 10 15 <210> SEQ ID NO 171 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 171 Gly Asp Arg Leu Val Phe Leu Glu Ala Leu Glu Arg Arg Ile Tyr 1 5 10 15 <210> SEQ ID NO 172 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 172 Leu Ala Tyr Arg Arg Arg Phe Pro Ala Val Ile Thr Arg Val Leu 1 5 10 15 <210> SEQ ID NO 173 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 173 Arg Arg Phe Pro Ala Val Ile Thr Arg Val Leu Pro Thr Arg Ile 1 5 10 15 <210> SEQ ID NO 174 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 174 Cys Ala Ile Ile His Ala Pro Ala Val Ser Gly Pro Gly Pro His 1 5 10 15 <210> SEQ ID NO 175 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 175 Pro Asn Gly Thr Arg Gly Phe Ala Pro Gly Ala Leu Arg Val Asp 1 5 10 15 <210> SEQ ID NO 176 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 176 Leu Arg Val Leu Arg Ala Ala Asp Gly Pro Glu Ala Cys Tyr Val 1 5 10 15 <210> SEQ ID NO 177 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 177 Asn Pro Arg Thr Ala Pro Arg Ser Leu 1 5 <210> SEQ ID NO 178 <211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic peptide <400> SEQUENCE: 178 Gly Pro Ala Asp Arg Arg Phe Val Ala Leu 1 5 10 <210> SEQ ID NO 179 <211> LENGTH: 2100 <212> TYPE: DNA <213> ORGANISM: HSV-2 <400> SEQUENCE: 179 atgcacgcca tcgctcccag gttgcttctt ctttttgttc tttctggtct tccggggaca 60 cgcggcgggt cgggtgtccc cggaccaatt aatcccccca acagcgatgt tgttttcccg 120 ggaggttccc ccgtggctca atattgttat gcctatcccc ggttggacga tcccgggccc 180 ttgggttccg cggacgccgg gcggcaagac ctgccccggc gcgtcgtccg tcacgagccc 240 ctgggccgct cgttcctcac gggggggctg gttttgctgg cgccgccggt acgcggattt 300 ggcgcaccca acgcaacgta tgcggcccgt gtgacgtact accggctcac ccgcgcctgc 360 cgtcagccca tcctccttcg gcagtatgga gggtgtcgcg gcggcgagcc gccgtcccca 420 aagacgtgcg ggtcgtacac gtacacgtac cagggcggcg ggcctccgac ccggtacgct 480 ctcgtaaatg cttccctgct ggtgccgatc tgggaccgcg ccgcggagac attcgagtac 540 cagatcgaac tcggcggcga gctgcacgtg ggtctgttgt gggtagaggt gggcggggag 600 ggccccggcc ccaccgcccc cccacaggcg gcgcgtgcgg agggcggccc gtgcgtcccc 660 ccggtccccg cgggccgccc gtggcgctcg gtgcccccgg tatggtattc cgcccccaac 720 cccgggtttc gtggcctgcg tttccgggag cgctgtctgc ccccacagac gcccgccgcc 780 cccagcgacc taccacgcgt cgcttttgct ccccagagcc tgctggtggg gattacgggc 840 cgcacgttta ttcggatggc acgacccacg gaagacgtcg gggtcctgcc gccccattgg 900 gcccccgggg ccctagatga cggtccgtac gcccccttcc caccccgccc gcggtttcga 960 cgcgccctgc ggacagaccc cgagggggtc gaccccgacg ttcgggcccc ccgaaccggg 1020 cggcgcctca tggccttgac cgaggacacg tcctccgatt cgcctacgtc cgctccggag 1080 aagacgcccc tccctgtgtc ggccaccgcc atggcaccct cagtcgaccc aagcgcggaa 1140 ccgaccgccc ccgcaaccac tactcccccc gacgagatgg ccacacaagc cgcaacggtc 1200 gccgttacgc cggaggaaac ggcagtcgcc tccccgcccg cgactgcatc cgtggagtcg 1260 tcgccactcc ccgccgcggc ggcggcaacg cccggggccg ggcacacgaa caccagcagc 1320 gcctccgcag cgaaaacgcc ccccaccaca ccagccccca cgaccccccc gcccacgtct 1380 acccacgcga ccccccgccc cacgactccg gggccccaaa caacccctcc cggacccgca 1440 accccgggtc cggtgggcgc ctccgccgcg cccacggccg attcccccct caccgcctcg 1500 ccccccgcta ccgcgccggg gccctcggcc gccaacgttt cggtcgccgc gaccaccgcc 1560 acgcccggaa cccggggcac cgcccgtacc cccccaacgg acccaaagac gcacccacac 1620 ggacccgcgg acgctccccc cggctcgcca gcccccccac cccccgaaca tcgcggcgga 1680 cccgaggagt ttgagggcgc cggggacggc gaaccccccg aggacgacga cagcgccacc 1740 ggcctcgcct tccgaactcc gaaccccaac aaaccacccc ccgcgcgccc cgggcccatc 1800 cgccccacgc tcccgccagg aattcttggg ccgctcgccc ccaacacgcc tcgccccccc 1860 gcccaagctc ccgctaagga catgccctcg ggccccacac cccaacacat ccccctgttc 1920 tggttcctaa cggcctcccc tgctctagat atcctcttta tcatcagcac caccatccac 1980 acggcggcgt tcgtttgtct ggtcgccttg gcagcacaac tttggcgcgg ccgggcgggg 2040 cgcaggcgat acgcgcaccc gagcgtgcgt tacgtatgtc tgccacccga gcgggattag 2100 <210> SEQ ID NO 180 <211> LENGTH: 471 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 180 atgcagcatc accaccatca ccacctgggt ctggctgaca cggtggtcgc gtgcgtggcc 60 ctggccgcgt ttgacggcgg gtcgacggcc cccgaggtgg gcacgtacac ccccctgcgc 120 tacgcgtgcg tcctccgcgc gacccagccc ctgtacgcgc ggaccacccc cgccaaattt 180 tgggcggacg tgcgcgccgc cgcggaacac gtggaccttc gccccgcgtc ctcggcgccc 240 cgggcgcccg tgagcgggac ggcagacccc gccttcctgc tcgaagacct ggcggccttc 300 ccccccgccc ccctgaatag cgagtccgtg ctggggccgc gggtccgcgt cgtggacatc 360 atggcgcagt ttcggaaact gctcatgggc gacgaggaga ccgccgccct ccgggcgcac 420 gtgtccggga ggcgcgcgac cgggctgggc ggcccgccac gcccatagtg a 471 <210> SEQ ID NO 181 <211> LENGTH: 155 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 181 Met Gln His His His His His His Leu Gly Leu Ala Asp Thr Val Val 5 10 15 Ala Cys Val Ala Leu Ala Ala Phe Asp Gly Gly Ser Thr Ala Pro Glu 20 25 30 Val Gly Thr Tyr Thr Pro Leu Arg Tyr Ala Cys Val Leu Arg Ala Thr 35 40 45 Gln Pro Leu Tyr Ala Arg Thr Thr Pro Ala Lys Phe Trp Ala Asp Val 50 55 60 Arg Ala Ala Ala Glu His Val Asp Leu Arg Pro Ala Ser Ser Ala Pro 65 70 75 80 Arg Ala Pro Val Ser Gly Thr Ala Asp Pro Ala Phe Leu Leu Glu Asp 85 90 95 Leu Ala Ala Phe Pro Pro Ala Pro Leu Asn Ser Glu Ser Val Leu Gly 100 105 110 Pro Arg Val Arg Val Val Asp Ile Met Ala Gln Phe Arg Lys Leu Leu 115 120 125 Met Gly Asp Glu Glu Thr Ala Ala Leu Arg Ala His Val Ser Gly Arg 130 135 140 Arg Ala Thr Gly Leu Gly Gly Pro Pro Arg Pro 145 150 155 <210> SEQ ID NO 182 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 182 ctgggtctgg ctgacacggt ggtcgcgtgc gtg 33 <210> SEQ ID NO 183 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: PCR primer <400> SEQUENCE: 183 ccgttagaat tcactatggg cgtggcgggc c 31 

What is claimed:
 1. An isolated polypeptide comprising at least an immunogenic portion of an HSV antigen, wherein said antigen comprises an amino acid sequence set forth in any one of SEQ ID NO: 2, 3, 5, 6, 7, 10-12, 14-15, 17-18, 20-23, 25-33, 39-47, 50-51, 54-64, 74-75, 90-97, 120-121, 122-140, 142-143, 153-178, and
 181. 2. An isolated polynucleotide encoding a polypeptide of claim
 1. 3. An isolated polynucleotide of claim 2, wherein said polynucleotide comprises a sequence set forth in any one of SEQ ID NO: 1, 4, 8-9, 13, 16, 19 24, 35-38, 48-49, 52-53, 65-73, 76-89, 98-117, 118-119, 141, 144-152, 179-180 and 182-183.
 4. An isolated polypeptide comprising at least an immunogenic portion of a HSV UL46 antigen, wherein said antigen comprises an amino acid sequence set forth in SEQ ID NO: 15, and wherein said immunogenic portion is selected from the group consisting of SEQ ID NO: 27-33 and 59-62.
 5. An isolated polypeptide comprising at least an immunogenic portion of a HSV UL15 antigen, wherein said antigen comprises an amino acid sequence set forth in SEQ ID NO: 26, and wherein said immunogenic portion is selected from the group consisting of SEQ ID NO: 56-57.
 6. An isolated polypeptide comprising at least an immunogenic portion of a HSV US3 antigen, wherein said antigen comprises an amino acid sequence set forth in SEQ ID NO: 12, and wherein said immunogenic portion comprises SEQ ID NO:
 63. 7. An isolated polypeptide comprising at least an immunogenic portion of a HSV US8A antigen, wherein said antigen comprises an amino acid sequence set forth in SEQ ID NO: 7, and wherein said immunogenic portion comprises SEQ ID NO:
 64. 8. An isolated polypeptide comprising at least an immunogenic portion of a HSV2 UL39 antigen, wherein said antigen comprises an amino acid sequence set forth in SEQ ID NO:3, and wherein said immunogenic portion is selected from the group consisting of SEQ ID NOs:74-75.
 9. An isolated polypeptide comprising at least an immunogenic portion of a HSV2 ICP0 antigen, wherein said antigen comprises an amino acid sequence set forth in SEQ ID NOs:47, and wherein said immunogenic portion is selected from the group consisting of SEQ ID NO:46.
 10. A fusion protein comprising a polypeptide according to claim 1 and a fusion partner.
 11. A fusion protein according to claim 10, wherein the fusion partner comprises an expression enhancer that increases expression of the fusion protein in a host cell transfected with a polynucleotide encoding the fusion protein.
 12. A fusion protein according to claim 10, wherein the fusion partner comprises a T helper epitope that is not present within the polypeptide of claim
 1. 13. A fusion protein according to claim 10, wherein the fusion partner comprises an affinity tag.
 14. An isolated polynucleotide encoding a fusion protein according to claim
 10. 15. An isolated monoclonal or polyclonal antibody, or antigen-binding fragment thereof, that specifically binds to a polypeptide of claim
 1. 16. A pharmaceutical composition comprising a polypeptide according to claim 1 or a polynucleotide encoding said polypeptide, and a physiologically acceptable carrier.
 17. A pharmaceutical composition comprising a polypeptide according to claim 1, or a polynucleotide encoding said polypeptide, and an immunostimulant.
 18. The pharmaceutical composition of claim 17, wherein the immunostimulant is selected from the group consisting of a monophosphoryl lipid A, aminoalkyl glucosaminide phosphate, saponin, or a combination thereof.
 19. A method for stimulating an immune response in a patient, comprising administering to a patient a pharmaceutical composition according to any one of claims 16-18.
 20. A method for detecting HSV infection in a patient, comprising: (a) obtaining a biological sample from the patient; (b) contacting the sample with a polypeptide according to claim 1; and (c) detecting the presence of antibodies that bind to the polypeptide.
 21. The method according to claim 20, wherein the biological sample is selected from the group consisting of whole blood, serum, plasma, saliva, cerebrospinal fluid and urine.
 22. A method for detecting HSV infection in a biological sample, comprising: (a) contacting the biological sample with a binding agent which is capable of binding to a polypeptide according to claim 1; and (b) detecting in the sample a polypeptide that binds to the binding agent, thereby detecting HSV infection in the biological sample.
 23. The method of claim 22, wherein the binding agent is a monoclonal antibody.
 24. The method of claim 22, wherein the binding agent is a polyclonal antibody.
 25. The method of claim 22 wherein the biological sample is selected from the group consisting of whole blood, sputum, serum, plasma, saliva, cerebrospinal fluid and urine.
 26. A diagnostic kit comprising a component selected from the group consisting of: (a) a polypeptide according to claim 1; (b) a fusion protein according to claim 10; (c) at least one antibody, or antigen-binding fragment thereof, according to claim 15; and (d) a detection reagent.
 27. The kit according to claim 26, wherein the polypeptide is immobilized on a solid support.
 28. The kit according to claim 26, wherein the detection reagent comprises a reporter group conjugated to a binding agent.
 29. The kit of claim 28, wherein the binding agent is selected from the group consisting of anti-immunoglobulins, Protein G, Protein A and lectins.
 30. The kit of claim 28, wherein the reporter group is selected from the group consisting of radioisotopes, fluorescent groups, luminescent groups, enzymes, biotin and dye particles. 